Using Exhaled Breath Condensate, Aerosols and Gases for Detecting Biomarkers

ABSTRACT

An apparatus for detecting a biomarker includes a particulate capturing structure for receiving and capturing exhaled breath aerosol (EBA) particulate from airway linings of a user, the particulate capturing structure having an aerosol particulate testing system for receiving the captured particulate and detecting a first biomarker, wherein the aerosol particulate testing system includes a dissolvable EBA sample collector film for capturing EBA particulate. The apparatus may include a droplet harvesting structure for converting breath vapor to a fluid droplet for forming a fluid sample and a testing system having a biomarker testing zone for receiving the fluid sample and detecting a biomarker.

RELATED APPLICATIONS

This application relates to co-pending US Utility Patent ApplicationTitled: Using Exhaled Breath Condensate for Testing for a Biomarker ofCOVID-19, Ser. No. 16/876,054, filed 17 May 2020, and claims priority ofUS Provisional Applications Titled: A Low Cost, Scalable, Accurate, andEasy-to-Use Testing System for COVID-19, Ser. No. 63/012,247 filed 19Apr. 2020; Using Exhaled Breath Condensate for Testing for a Biomarkerof COVID-19, Ser. No. 63/019,378 filed 3 May 2020; and Using ExhaledBreath Condensate for Testing for a Biomarker of COVID-19, Ser. No.63/026,052 filed 17 May 2020; the disclosures of which are hereinincorporated by reference in their entireties.

TECHNICAL FIELD

The exemplary and non-limiting embodiments of this invention relategenerally to digital therapeutic systems, methods, devices and computerprograms and, more specifically, relate to digital therapeutic wearableelectronic garments for detecting a biomarker of a biological agent suchas a coronavirus.

The present invention also pertains to a device architecture,specific-use applications, and computer algorithms used with wearableelectronics with the capability to detect biometric parameters for thetreatment and monitoring of physiological conditions in humans andanimals.

BACKGROUND

This section is intended to provide a background or context to theexemplary embodiments of the invention as recited in the claims. Thedescription herein may include concepts that could be pursued but arenot necessarily ones that have been previously conceived, implemented ordescribed. Therefore, unless otherwise indicated herein, what isdescribed in this section is not prior art to the description and claimsin this application and is not admitted to be prior art by inclusion inthis section.

Governments around the world have instituted stay at home policies andthe lockdown of citizens to slow the spread of the COVID-19 virus. Thereare currently billions of people around the world that have halted theirusual employment, entertainment and socializing activities. Testing forbiomarkers that indicate exposure, infection and recovery from COVID-19can be used to enable a safer and more efficient restart of economicactivities, while minimizing the spread of the virus. For example, RNAtesting for active virus shows who is currently contagious. Antibodytesting can be used to find the members of a population that haverecovered from the virus and now may be immune to reinfection. Thisknowledge could enable precision social distancing and more effectivecontact tracing, with the re-employment of a growing workforce ofprotected individuals and consumers. Those who remain at-risk ofinfection and transmission can be kept sequestered until a vaccine orother solution such as a high success rate pharmaceutical therapy isdeveloped.

In immunochromagtography, a capture antibody is disposed onto a surfaceof a porous membrane, and a sample passes along the membrane. Analyte inthe sample is bound by the antibody which is coupled to a detectorreagent. As the sample passes through the area where the captureantibody is disposed, an analyte detector reagent complex is trapped,and a color develops that is proportional to the analyte present in thesample.

In a lateral flow assay, a liquid sample containing a target analyte(s)flows through a multi-zone transfer medium through capillary action. Thezones are typically made of polymeric strips enabling molecules attachedto the strips to interact with the target analyte. Usually, overlappingmembranes are mounted on a backing card to improve stability andhandling. The sample containing the target analyte and otherconstituents is ultimately received at an adsorbent sample pad whichpromotes wicking of the fluid sample through the multi-zone transfermedium.

The fluid sample is first received at a sample pad which may have buffersalts and surfactants disposed on or impregnated into it to improve theflow of the fluid sample and the interaction of the target analyte withthe various parts of the detection system. This ensures that the targetanalyte will bind to capture reagents as the fluid sample flows throughthe membranes. The treated sample migrates from the sample pad through aconjugate release pad. The conjugate release pad contains labeledantibodies that are specific to the target analyte and are conjugated tocolored or fluorescent indicator particles. The indicator particles aretypically, colloidal gold or latex microspheres.

At the conjugate release pad, the labeled antibodies, indicatorparticles and target analyte bind to form a target analyte-labeledantibody complex. If an analyte is present, the fluid sample nowcontains the indicator particles conjugated to the labeled antibody andbound to the target analyte (i.e., the target analyte-labeled antibodycomplex) along with separate labeled antibodies conjugated to theindicator particles that have not been bound to the target analyte. Thefluid sample migrates along the strip into a detection zone.

The detection zone is typically a nitrocellulose porous membrane and hasspecific biological components (usually antibodies or antigens) disposedon or impregnated in it forming a test line zone(s) and control linezone. The biological components react with the target analyte-labeledantibody complex. For example, the target analyte-labeled antibodycomplex will bind to a specifically selected primary antibody that isdisposed at the test line through competitive binding. This results incolored or fluorescent indicator particles accumulating at the test linezone making a detectable test line that indicates the target analyte ispresent in the fluid sample.

The primary antibody does not bind to the separate labeled antibodiesand they continue to flow along with the fluid sample. At a control linezone, a secondary antibody binds with the separate labeled antibodiesconjugated to the indicator particles and thereby indicates the properliquid flow through the strip.

The fluid sample flows through the multi-zone transfer medium of thetesting device through the capillary force of the materials making upthe zones. To maintain this movement, an absorbent pad is attached asthe end zone of the multi-zone transfer medium. The role of theabsorbent pad is to wick the excess reagents and prevent back-flow ofthe fluid sample.

The constituents are selected and disposed on the membranes so that ifthere is no target analyte present in the fluid sample, there will be notarget analyte-labeled antibody complex present that flows through thetest line zone. In this case there will be no accumulation of thecolored or fluorescent particles and no detectable test line will form.Even if there is no analyte and thus no test line, there will still be acontrol line formed because the secondary antibody still binds to theseparate labeled antibodies that flow along with the fluid sample.

The test and control lines may appear with different intensities anddepending on the device structure and the indicator particles can beassessed by eye or using an optical or other electronic reader. Multipleanalytes can be tested simultaneously under the same conditions withadditional test line zones of antibodies specific to different analytesdisposed in the detection zone in an array format. Also, multiple testline zones loaded with the same antibody can be used for quantitativedetection of the target analyte. This is often called a ‘ladder bars’assay based on the stepwise capture of colorimetric conjugate-antigencomplexes by the immobilized antibody on each successive line. Thenumber of lines appearing on the strip is directly proportional to theconcentration of the target analyte.

What is needed now is a low cost, scalable, accurate and easy-to-usetesting system that can be deployed to the masses via the mail orcourier for at-home use.

SUMMARY

The below summary section is intended to be merely exemplary andnon-limiting. The foregoing and other problems are overcome, and otheradvantages are realized, by the use of the exemplary embodiments of thisinvention.

In accordance with an aspect of the invention, an apparatus fordetecting a biomarker includes a particulate capturing structure forreceiving and capturing exhaled breath aerosol (EBA) particulate fromairway linings of a user, the particulate capturing structure having anaerosol particulate testing system for receiving the capturedparticulate and detecting a first biomarker, wherein the aerosolparticulate testing system includes a dissolvable EBA sample collectorfilm for capturing EBA particulate. The dissolvable EBA sample collectorfilm includes a first reagent for reacting with at least one constituentof the captured particulate in a detection reaction for detecting thefirst biomarker. The detection reaction generates at least one of achange in an optical signal and an electrical signal dependent on thefirst biomarker. The first reagent is bound to a first nanoparticle andheld in place at the insoluble testing area. The EBA particulateincludes non-soluble particulates and droplet particulates, and thedissolvable EBA collector film includes a tacky surface for adhering toand capturing the non-soluble particulates and water soluble bulk forcapturing droplet particulates.

In accordance with another aspect of the invention an apparatuscomprises at least one processor, at least one memory including computerprogram code, the at least one memory and the computer program codeconfigured to, with the at least one processor, cause the apparatus toperform at least the following: detecting one or more biometricparameters using a particulate capturing structure for receiving andcapturing exhaled breath aerosol (EBA) particulate from airway liningsof a user, the particulate capturing structure having an aerosolparticulate testing system for receiving the captured particulate anddetecting a first biomarker, wherein the aerosol particulate testingsystem includes a dissolvable EBA sample collector film for capturingEBA particulate, where the biometric parameters are biomarkers dependenton at least one physiological change to a patient in response to aconcerning condition such as a virus infection; receiving the one ormore biometric parameters and applying probabilistic analysis todetermine if at least one physiological change threshold has beenexceeded dependent on the probabilistic analysis of the one ore morebiometric parameters; and activating an action depending on thedetermined exceeded said at least one physiological change. The one ormore biometric parameters can be further detected using a dropletharvesting structure for converting breath vapor to a fluid droplet forforming a fluid sample and a testing system having a biomarker testingzone for receiving the fluid sample and detecting the biometricparameter; and wherein the probabilistic analysis is applied to the oneor more biometric parameters to determine if the at least onephysiological change threshold has been exceeded dependent on theprobabilistic analysis of the one ore more biometric parameters detectedfrom both the captured particulates and the fluid sample.

BRIEF DESCRIPTION OF THE DRAWINGS

The foregoing and other aspects of exemplary embodiments of thisinvention are made more evident in the following Detailed Description,when read in conjunction with the attached Drawing Figures, wherein:

FIG. 1 shows a Lateral Flow Assay (LFA) testing system showing ananalyte sample added to a sample pad;

FIG. 2 shows the LFA with an analyte-labeled antibody complex formed ata conjugate release pad;

FIG. 3 shows the binding of analyte at a test line indicating thepresence of the analyte;

FIG. 4 shows the mechanism of a bioreceptor detection system;

FIG. 5 is a side view of a wearable electronic breath chemistry sensor;

FIG. 6 is a top view of a wearable electronic breath chemistry sensor;

FIG. 7 is an isolated view of a Exhaled Breath Condensate (EBC) dropletsample collector;

FIG. 8 is a top view showing a step for forming the EBC dropletcollector;

FIG. 9 is a top view showing another step for forming the EBC dropletsample collector;

FIG. 10 is a top view showing still another step for forming the EBCdroplet sample collector;

FIG. 11 is a top view showing yet another step for forming the EBCdroplet sample collector;

FIG. 12 illustrates the EBC sample collector showing EBS droplets;

FIG. 13 illustrates the EBC sample collector applied to an LFA testingsystem;

FIG. 14 is an exploded view showing the screen printed hydrophilicchannels, screen printed hydrophobic field and thermal mass substrate ofthe EBC sample collector;

FIG. 15 is an exploded view showing the constituent elements of a LFA;

FIG. 16 illustrates an embodiment of the LFA including photonicemitter/detector electronics;

FIG. 17 illustrates the EBC sample collector applied to a nanoscalebioreceptor testing system and showing a pull tab for holding backcollected droplets on the sample pad;

FIG. 18 is a perspective view showing the EBC sample collector appliedto a testing system;

FIG. 19 is an isolated view showing the pull table disposed between thesample pad and conjugate release pad;

FIG. 20 is an isolated view of a screen printed EBC sample collectorwith a fluid transfer aperture;

FIG. 21 is a cross-section view showing a fluid sample collected fromthe EBC sample collector flowing between a photonics emitter/detectorpair;

FIG. 22 shows the side views of the steps for building up an LFA testingsystem;

FIG. 23 shows the top view of the steps for building up an LFA testingsystem;

FIG. 24 shows a 4×9 ganged multiple-up sheet of LFA testing systemsformed as a batch;

FIG. 25 shows a roll-to-roll manufacturing process for forming a roll ofbottom adhesive/backing substrate/top adhesive;

FIG. 26 is a perspective view illustrating the bottom adhesive/backingsubstrate/top adhesive stack;

FIG. 27 shows a roll-to-roll manufacturing process for forming theconstituent elements of an LFA on a roll of bottom adhesive/backingsubstrate/top adhesive;

FIG. 28 shows the LFA testing system formed by the roll-to-roll processcut from a continuous roll and showing a section of top adhesive foradhering the LFA testing system to a separately formed ENC samplecollector;

FIG. 29 shows the LFA testing system formed by the roll-to-roll processcut from a continuous roll and showing a section of bottom adhesive forsticking onto a wearable garment such as a face mask;

FIG. 30 shows a sheet of substrate with a hydrophobic field coating on athermal mass substrate with droplet collection holes;

FIG. 31 shows the sheet of substrate with the hydrophobic field coatingon the thermal mass substrate with droplet collection holes having acoating of hydrophilic channels;

FIG. 32 shows the EBC sample collector and testing system withelectronics for wireless data acquisition and transmission along withseparate trusted receiver and public blockchain data path and storage;

FIG. 33 shows the manufacturing processes for a heat bonded face mask;

FIG. 34 shows the fabric, filter and other layers bonded through aroll-to-roll lamination processor individually cut into blanks forforming a pre-form mask stack;

FIG. 35 shows other materials such as biological reactive silver fabricand hot melt adhesive of the pre-form mask stack;

FIG. 36 is an exploded view of a mask stack;

FIG. 37 shows the fold lines of the mask stack for first and second heatpress operations;

FIG. 38 shows the folded, pressed and heat bonded mask;

FIG. 39 shows the attachment of the EBC collector and testing system tothe folded mask;

FIG. 40 shows the step of turning the folded mask inside out to disposethe EBC collector and testing system on the inside of the mask;

FIG. 41 shows a heat press operation to bond elastic straps onto thefolded mask;

FIG. 42 shows the mask with the EBC collector and testing systemdisposed inside the mask within the concentrated atmosphere of exhaledbreath;

FIG. 43 shows a conventional bendable metal nose seal that is disposedwithin the folds of the mask at a location corresponding to the bridgeof a user's nose;

FIG. 44 shows a replaceable adhesive nose strip that is disposed on theoutside of the folds of the mask at a location corresponding to thebridge of a user's nose;

FIG. 45 shows the components of a magnetic removable nose seal;

FIG. 46 is an exploded view of a testing system including a dissolvableflow dam that holds back collected EBC on the sample pad until enoughhas been accumulated to be released onto the conjugate release pad andflush the fluid sample through the components of the testing system;

FIG. 47 is an isolated view showing the dissolvable flow dam insertedbetween the sample pad and the conjugate release pad;

FIG. 48 is an isolated view showing after the dissolvable flow dam hasbeen dissolved away to release the accumulated fluid sample from thesample pad to the conjugate release pad;

FIG. 49 is an isolated view showing a dissolvable EBC droplet and EBAparticulate collector;

FIG. 50 is a cross section side view showing a section of thedissolvable droplet and particulate collector having particulate anddroplets impinged on the surface;

FIG. 51 is a cross section side view showing the section of thedissolvable droplet and particulate collector having particulateembedded into the dissolvable capture film and droplets dissolved intoand causing a detection reaction with a detection reagent of thedissolvable capture film;

FIG. 52 is a top view showing the inventive testing system including adissolvable EBC droplet and EBA particulate collector having capturedaerosol droplets and aerosol particulate;

FIG. 53 is an isolated perspective view showing the dissolvable EBCdroplet and EBA particulate collector having captured aerosol dropletsand aerosol particulate;

FIG. 54 is a top view showing the inventive testing system including adissolvable EBC droplet and EBA particulate collector before capturingaerosol droplets and aerosol particulate;

FIG. 55 is a top view showing the inventive testing system including adissolvable EBC droplet and EBA particulate collector after capturingaerosol droplets and aerosol particulate;

FIG. 56 is a top view showing the inventive testing system including adissolvable EBC droplet and EBA particulate collector installed onto aface mask substrate along with a plurality of gas sensors for detectingvolatile and gas constituents of the exhaled breath and/or ambientatmosphere;

FIG. 57 is a cross section side view showing a section of thedissolvable droplet and particulate collector having particulate anddroplets impinged on the surface placed in a beaker of dissolvingliquid;

FIG. 58 is a cross section side view showing a section of thedissolvable droplet and particulate collector having the particulatereleased into and the droplets dissolved into the beaker of dissolvingliquid;

FIG. 59 is a block diagram of one possible and non-limiting exemplarysystem in which the exemplary embodiments may be practiced;

FIG. 60 is a logic flow diagram for Applied Probabilistic Analysis toDetermine COVID-19 Exposure, and illustrates the operation of anexemplary method, a result of execution of computer program instructionsembodied on a computer readable memory, functions performed by logicimplemented in hardware, and/or interconnected means for performingfunctions in accordance with exemplary embodiments;

FIG. 61 is a logic flow diagrams for Data Acquisition and Transmissionfor Trusted Receiver and Contract Tracing Uses, and illustrate theoperation of an exemplary method, a result of execution of computerprogram instructions embodied on a computer readable memory, functionsperformed by logic implemented in hardware, and/or interconnected meansfor performing functions in accordance with exemplary embodiments;

FIG. 62 is a perspective view of an embodiment of a EBC/EBA collectionsystem;

FIG. 63 is a perspective view of the EBC/EBA collection system showing apipette and pipette guide;

FIG. 64 is an exploded view showing the constituent parts of theembodiment of the EBC/EBA collection system;

FIG. 65 is another exploded view showing the constituent parts of theEBC/EBA collection system;

FIG. 66 is a cross-sectional view of the EBC/EBA collection system;

FIG. 67 illustrates the use of the EBC/EBA collection system forobtaining biomarker samples from the lungs of a user;

FIG. 68 is an isolated view showing the mouthpiece, cap, base,dissolvable EBA sample collector and inner cylinder of the embodiment ofthe EBC/EBA collection system;

FIG. 69 is an isolated view showing the dissolvable EBA sample collectorand inner cylinder having captured EBA particles and droplets;

FIG. 70 shows the inner cylinder submersed in a solvent for dissolvingthe dissolvable EBA sample collector to acquire the captured EBAparticles and droplets for biomarker testing;

FIG. 71 is an isolated view of a section of an embodiment of thedissolvable EBA sample collector forming an aerosol particulate testingsystem having captured EBA particulate, insoluble testing areas anddissolvable capture film areas; and

FIG. 72 shows a series of side views of the embodiment of thedissolvable EBS sample collector capturing EBA droplets and/orparticulate showing the aerosol particulate testing system with targetanalytes captured and bound to the insoluble testing areas;

FIG. 73 shows nanoparticles held in a trench in a substrate where thenanoparticles include capture antibodies or other reagent fixed to them;

FIG. 74 shows the EBA particles and droplets being rinsed from thedissolvable EBA sample collector to form a fluid sample that includesany biomarker analytes contained in the particles or droplets; and

FIG. 75 illustrates the EBA/EBC testing system with a wirelesscommunication electronic circuit that detects a result of the testingfor at least one of the first and second biomarker and communicating theresult to a wireless receiver.

DETAILED DESCRIPTION

Below are provided further descriptions of various non-limiting,exemplary embodiments. The exemplary embodiments of the invention, suchas those described immediately below, may be implemented, practiced orutilized in any combination (e.g., any combination that is suitable,practicable and/or feasible) and are not limited only to thosecombinations described herein and/or included in the appended claims.

The word “exemplary” is used herein to mean “serving as an example,instance, or illustration.” Any embodiment described herein as“exemplary” is not necessarily to be construed as preferred oradvantageous over other embodiments. All of the embodiments described inthis Detailed Description are exemplary embodiments provided to enablepersons skilled in the art to make or use the invention and not to limitthe scope of the invention which is defined by the claims.

Many configurations, embodiments, methods of manufacture, algorithms,electronic circuits, microprocessor, memory and computer softwareproduct combinations, networking strategies, database structures anduses, and other aspects are disclosed herein for a wearable electronicdigital therapeutic device and system that has a number of medical andnon-medical uses.

Researchers have been able to detect biomarkers in the breath ofpatients that have interstitial lung disease (see, Hayton, C.,Terrington, D., Wilson, A. M. et al. Breath biomarkers in idiopathicpulmonary fibrosis: a systematic review. Respir Res 20, 7 (2019).https://doi.org/10.1186/s12931-019-0971-8). An embodiment of theinventive testing system detects COVID-19 specific biomarkers present inthe breath of infected, infectious or post-recovery individuals.

The inventive COVID-19 testing system has the ability to coalesce breathvapor into droplets and then pass the droplet sample over a fluidicbiosensor, such as a Lateral Flow Assay (LFA) or Nanoscale-Bioreceptor(NS-BR) to enable a very low cost, manufacturable at-scale testingsystem that can be distributed to the masses for at-home triage testing.The inventive testing system can also be used for other biometric andenvironmental testing applications other than for virus detection.

LFAs can be used for the detection of a wide range of biomarkers presentin the breath including cytokines, proteins, haptens (elicit theproduction of antibodies), nucleic acids and amplicons (pieces of RNAand DNA) (SEE, Corstjens P L, de Dood C J, van der Ploeg-van Schip J J,et al. Lateral flow assay for simultaneous detection of cellular- andhumoral immune responses. Clin Biochem. 2011; 44(14-15):1241-1246.doi:10.1016/j.clinbiochem.2011.06.983).

The NSF Center for High-Rate Nanomanufacturing (CHN) has a directedassembly technique for high throughput manufacturing of NS-BRs. Thetechnique is proven to selectively assemble nanoparticles coated withspecific antibodies onto a single microchip surface for the simultaneousdetection of multiple biomarkers. Early results suggested sensitivity toconcentrations of much less than 1 ng/mL—a large increase in sensitivityrelative to that of the commercially available ELISA detection kit. TheCHN biosensor is very small, about 0.25 mm in diameter, and hasadvantages compared to traditional in vitro techniques because itenables disease markers detection with less false positives with a verylow detection limit. This capability will be very useful for detectingvery small changes in biomarker concentration in disease monitoring(see, Highly sensitive microscale in vivo sensor enabled byelectrophoretic assembly of nanoparticles for multiple biomarkerdetection, Malima et al., Lab Chip, 2012, 12, 4748-4754).

Exhaled breath collection has long been recognized as requiring theleast invasive methods, and so is preferred for environmental and publichealth studies. In contrast to blood and urine, breath sampling does notrequire trained medical personnel or privacy, does not createpotentially infectious wastes, and can be done essentially anywhere inany time frame. Although the Exhaled Breath Condensate (EBC) formatdiscriminates against most non-polar VOCs, it has the advantage ofcollecting polar compounds and heavier analytes including semi- andnon-volatile organics, cytokines, proteins, cellular fragments, DNA, andbacteria. Exhaled breath also contains tiny aerosols (including bothliquid and solid particles) that are created by surface film disruptionat the alveolar level and by upper airway turbulence. These aerosolsgive mobility to materials that are otherwise relegated to the liquidlayers within the lung and, as such, are that part of the EBC whichcontributes the non-volatile analytes. Exhaled Breath Aerosols (EBA)impacts surfaces forming a layer; only the residues are subsequentlyharvested using various wiping or extraction techniques. Sampling issimplified as the subject only wears a mask for some period of time.This could be either a surgical-style mask for public health monitoring,or a standard mask used for occupational respiratory protection (FIG.1). The elegance of the technique is that only dried residues aretransported to the lab; there are no special requirements for shipping(see, Pleil J D, Wallace M A G, Madden M C. Exhaled breath aerosol(EBA): the simplest non-invasive medium for public health andoccupational exposure biomonitoring. J Breath Res. 2018; 12(2):027110.Published 2018 Feb. 6. doi:10.1088/1752-7163/aa9855).

The usual methods for obtaining clinical specimens from the respiratorytract are nasopharyngeal or oropharyngeal swabs, nasopharyngealaspirates and nasal washes, tracheal aspirates, bronchoalveolar lavage,or the collection of sputum. Each of these techniques has drawbacks:Nasopharyngeal and oropharyngeal swabs, aspirates, and washes providemucus from the upper respiratory tract, which does not always containthe same viral load or the same species of viruses as the lowerrespiratory tract. The collection of aerosol particles produced bypatients during coughing and tidal breathing potentially provides anon-invasive method for the collection of diagnostic specimens ofrespiratory viruses. Respiratory viruses have been detected in theexhaled breath and cough aerosols from infected patients, especiallyinfluenza virus. Microbial aerosols may also be more representative oflower respiratory tract disease in viral illnesses in which sputumproduction is not common. Because exhaled aerosol collection isnon-invasive, repeated sample collection should be more acceptable topatients than traditional methods. If the limitations can be overcome,exhaled aerosol analysis could become a useful tool for the diagnosis ofrespiratory infections and for monitoring the course of illness andresponse to treatment (see, Fennelly K P, Acuna-Villaorduna C,Jones-Lopez E, Lindsley W G, Milton D K. Microbial Aerosols: NewDiagnostic Specimens for Pulmonary Infections. Chest. 2020;157(3):540-546. doi:10.1016/j.chest.2019.10.012).

Usually, a thermoelectric cooling module is required to collectsufficient EBC volume for analyses but researchers have shown thefeasibility of cytokine and chemokine detection in EBC collecteddirectly from the ventilator circuit without the use of a cooling modulefrom swivel-derived exhaled breath condensate (SEBC). SEBC was collectedfrom the swivel adapter in a ventilator and cytokines and chemokines inSEBC was detected with a multiplex immunoassay. Twenty-nine SEBC sampleswere obtained from 13 ICU patients. IL-1β, IL-4, IL-8, and IL-17 weredetected in more than 90% of SEBC samples, and significant correlationsbetween multiple cytokines and chemokines were found. Severalsignificant correlations were found between cytokines and chemokines inSEBC and mechanical ventilation parameters and serum lactateconcentrations. This pilot study showed that it is feasible to detectcytokines and chemokines in SEBC samples obtained without a coolingmodule (see, an der Zee P, van Walree I, Fijen J W, et al. Cytokines andChemokines Are Detectable in Swivel-Derived Exhaled Breath Condensate(SEBC): A Pilot Study in Mechanically Ventilated Patients. Dis Markers.2020; 2020:2696317. Published 2020 Jan. 11. doi:10.1155/2020/2696317).

There are more than 2000 compounds identified in EBC (see, Montuschi P,Mores N, Trové A, Mondino C, Barnes P J. The electronic nose inrespiratory medicine. Respiration. 2013; 85(1):72-84) and many of themare considered to represent sensitive biomarkers of lung diseases (see,Sapey E, editor. Bronchial Asthma: Emerging Therapeutic Strategies.Rijeka: InTech. Biomarkers present in EBC depict the processes occurringin lungs much more than those in the entire body system. Therefore,particular profiles of exhaled biomarkers can reveal informationexclusively applicable to lung disease diagnoses. EBC is a biologicalmatrix reflecting the composition of the bronchoalveolar extra-cellularlung fluid. The main advantage of EBC as of a matrix is its specificityfor the respiratory tract (the liquid is not influenced by processoccurring in other parts of the body) (see, Molecular Diagnostics ofPulmonary Diseases Based on Analysis of Exhaled Breath Condensate,Tereza Kačerová, Petr Novotny, Ján Boroň and Petr Kačer Submitted: Oct.9 2016 Reviewed: Jan. 25 2018 Published: Sep. 5 2018, DOI:10.5772/intechopen.7440).

The surfaces in all parts of the lung down to the alveoli are coatedwith an aqueous mucous layer that can be aerosolized and carry along avariety of non-volatile constituents. EBC and EBA are different types ofbreath matrices used to assess human health and disease state. EBArepresents a fraction of total EBC, and is targeted to larger molecules,such as fatty acids and cytokines, as well as cellular fractions,proteins, viruses, and bacteria instead of the gas-phase. There is awide variety of compounds, such as volatile organic compounds (VOCs),NO, CO2, NH3, cytokines, and hydrogen peroxide (H2O2) in exhaled breathcondensate (EBC), and exhaled breath aerosol (EBA). VOCs located infatty tissues are released to the blood and are then exchanged into thebreath through the alveoli and airways in the lungs. A portion of VOCsare also retained within the respiratory tract after exposure. Thus,breath concentrations of VOCs are representative of bloodconcentrations, but samples can be obtained non-invasively with littlediscomfort to the individual (see, Wallace M A G, Pleil J D. Evolutionof clinical and environmental health applications of exhaled breathresearch: Review of methods and instrumentation for gas-phase,condensate, and aerosols. Anal Chim Acta. 2018; 1024:18-38.doi:10.1016/j.aca.2018.01.069).

EBC and EBA are valuable non-invasive biological media used for thequantification of biomarkers. EBC contains exhaled water vapor, solublegas-phase (polar) organic compounds, ionic species, plus other speciesincluding semi- and non-volatile organic compounds, proteins, cellfragments, DNA, dissolved inorganic compounds, ions, and micro-biota(bacteria and viruses) dissolved in the co-collected EBA (see, intersBR, Pleil J D, Angrish M M, Stiegel M A, Risby T H, Madden M C.Standardization of the collection of exhaled breath condensate andexhaled breath aerosol using a feedback regulated sampling device. JBreath Res. 2017; 11(4):047107. Published 2017 Nov. 1.doi:10.1088/1752-7163/aa8bbc).

An earlier reference reports detecting influenza virus RNA in theexhaled breath of 4 (33%) subjects: three (60%) of the five patientsinfected with influenza A virus and one (14%) of the seven infected withinfluenza B virus. Although a sample of EBC may have virus RNA in lessconcentrations than a nasal swab, these tests did determine detectableinfluenza virus RNA in exhaled breath. Concentrations in exhaled breathsamples ranged from 48 to 300 influenza virus RNA copies per filter onthe positive samples, corresponding to exhaled breath generation ratesranging from 3.2 to 20 influenza virus RNA copies per minute. Theresearchers note possible explanations for not detecting influenza virusRNA in a larger proportion of subjects may be due to short samplecollection times, the large heterogeneity in the virus production amonginfected patients and the detection limit for our qPCR method (see,Fabian P, McDevitt J J, DeHaan W H, et al. Influenza virus in humanexhaled breath: an observational study. PLoS One. 2008; 3(7):e2691.Published 2008 Jul. 16. doi:10.1371/journal.pone.0002691). Thisreference shows that nasal and throat swabs will typically have more RNAconcentrations than EBC. However, the virus RNA is clearly present inEBC and a EBC testing system with enough sensitivity should be effectiveat detecting the virus RNA.

Electron Microscope (SEM), polymerase chain reaction (PCR) andcolorimetry (VITEK 2) for bacteria and viruses show that bacteria andviruses in EBC can be rapidly collected with an observed efficiency of100 mL EBC within 1 min (see, Xu Z, Shen F, Li X, Wu Y Chen Q, et al.(2012) Molecular and Microscopic Analysis of Bacteria and Viruses inExhaled Breath Collected Using a Simple Impaction and Condensing Method.PLoS ONE 7(7): e41137. doi:10.1371/journal.pone.0041137).

Exhaled breath contains volatile organic compounds (VOCs), a collectionof hundreds of small molecules linked to several physiological andpathophysiological processes. Analysis of exhaled breath throughgas-chromatography and mass-spectrometry (GC-MS) has resulted in anaccurate diagnosis of ARDS in several studies. Most identified markersare linked to lipid peroxidation. Octane is one of the few markers thatwas validated as a marker of ARDS and is pathophysiologically likely tobe increased in ARDS (see, Bos L D J. Diagnosis of acute respiratorydistress syndrome by exhaled breath analysis. Ann Transl Med. 2018;6(2):33. doi:10.21037/atm.2018.01.17).

FIG. 1 shows a Lateral Flow Assay (LFA) testing system showing ananalyte sample added to a sample pad. FIG. 2 shows the LFA with ananalyte-labeled antibody complex formed at a conjugate release pad. FIG.3 shows the binding of analyte at a test line indicating the presence ofthe analyte. In a lateral flow assay, a liquid sample containing atarget analyte(s) flows through a multi-zone transfer medium throughcapillary action. The zones are typically made of polymeric stripsenabling molecules attached to the strips to interact with the targetanalyte. Usually, overlapping membranes are mounted on a backing card toimprove stability and handling. The sample containing the target analyteand other constituents is ultimately received at an adsorbent sample padwhich promotes wicking of the fluid sample through the multi-zonetransfer medium.

The fluid sample is first received at a sample pad which may have buffersalts and surfactants disposed on or impregnated into it to improve theflow of the fluid sample and the interaction of the target analyte withthe various parts of the detection system. This ensures that the targetanalyte will bind to capture reagents as the fluid sample flows throughthe membranes. The treated sample migrates from the sample pad through aconjugate release pad. The conjugate release pad contains labeledantibodies that are specific to the target analyte and are conjugated tocolored or fluorescent indicator particles. The indicator particles aretypically, colloidal gold or latex microspheres.

At the conjugate release pad, the labeled antibodies, indicatorparticles and target analyte bind to form a target analyte-labeledantibody complex. If an analyte is present, the fluid sample nowcontains the indicator particles conjugated to the labeled antibody andbound to the target analyte (i.e., the target analyte-labeled antibodycomplex) along with separate labeled antibodies conjugated to theindicator particles that have not been bound to the target analyte. Thefluid sample migrates along the strip into a detection zone.

The detection zone is typically a nitrocellulose porous membrane and hasspecific biological components (usually antibodies or antigens) disposedon or impregnated in it forming a test line zone(s) and control linezone. The biological components react with the target analyte-labeledantibody complex. For example, the target analyte-labeled antibodycomplex will bind to a specifically selected primary antibody that isdisposed at the test line through competitive binding. This results incolored or fluorescent indicator particles accumulating at the test linezone making a detectable test line that indicates the target analyte ispresent in the fluid sample.

The primary antibody does not bind to the separate labeled antibodiesand they continue to flow along with the fluid sample. At a control linezone, a secondary antibody binds with the separate labeled antibodiesconjugated to the indicator particles and thereby indicates the properliquid flow through the strip.

The fluid sample flows through the multi-zone transfer medium of thetesting device through the capillary force of the materials making upthe zones. To maintain this movement, an absorbent pad is attached asthe end zone of the multi-zone transfer medium. The role of theabsorbent pad is to wick the excess reagents and prevent back-flow ofthe fluid sample.

The constituents are selected and disposed on the membranes so that ifthere is no target analyte present in the fluid sample, there will be notarget analyte-labeled antibody complex present that flows through thetest line zone. In this case there will be no accumulation of thecolored or fluorescent particles and no detectable test line will form.Even if there is no analyte and thus no test line, there will still be acontrol line formed because the secondary antibody still binds to theseparate labeled antibodies that flow along with the fluid sample.

The test and control lines may appear with different intensities anddepending on the device structure and the indicator particles can beassessed by eye or using an optical or other electronic reader. Multipleanalytes can be tested simultaneously under the same conditions withadditional test line zones of antibodies specific to different analytesdisposed in the detection zone in an array format. Also, multiple testline zones loaded with the same antibody can be used for quantitativedetection of the target analyte. This is often called a ‘ladder bars’assay based on the stepwise capture of colorimetric conjugate-antigencomplexes by the immobilized antibody on each successive line. Thenumber of lines appearing on the strip is directly proportional to theconcentration of the target analyte.

Another testing system that can be used with the inventive EBCcollection system uses a nano-scale bioreceptor (NS-BR). Similar to LFA,NS-BR has the potential of a much higher sensitivity and can be used toprovide a direct-to-electrical signal to enable, for example, easywireless connectivity. The inventive EBC collection system with NS-BRtesting is easily deployable as a compliment to existing Contact TracingAPPs. The nanoscale dimensions mean many detectors are made at once on asingle wafer for lower cost, high throughput manufacturing.

FIG. 4 shows the mechanism of a bioreceptor detection system.Simplistically, the main components of a fluidic biosensor includes asample source (a); a bioreceptor area that is functionalized with ananalyte-specific bioreceptor (b); and a transducer for generating areadable signal (c). The bioreceptor is matched to a specific targetbiomarker for lock and key selectivity screening. A fluid sample withsome concentration of the target biomarker analyte (possibly as small asa single molecule) flows onto the bioreceptor field. Some of thebioreceptor “locks” receive the biomarker “keys.” This causes adetectable change in the output of the transducer that transforms thebioreceptor output into a readable signal for amplification and dataprocessing.

For example, the desired biomarker analyte can also be an antibody thatindicates the recovery from a Covid-19 infection. A fluid sample can bereceived as a droplet of sweat or breath or other body fluid and if thetarget antibody is present in the sample it interacts with theanalyte-specific bioreceptor. The bioreceptor outputs a signal withdefined sensitivity and the transducer generates, for example, a changein an electrical characteristic such as conductivity, indicating thepresence of the antibody biomarker in the fluid sample.

In accordance with an embodiment, an apparatus for detecting a biomarkercomprises a droplet harvesting and channeling structure for convertingvapor to a fluid droplet and a fluidic biosensor including a samplesource having a biomarker analyte, a bioreceptor area functionalizedwith an analyte-specific bioreceptor, and a transducer for generating areadable signal depending on a change in the bioreceptor in response toreceiving the biomarker analyte from the sample source.

Using nano-scale sensor technology enables detection of very lowconcentrations of the target analyte(s) such as virus RNA and/orantibodies while avoiding the need for drawing blood. In accordance withan embodiment of the inventive testing system, a droplet harvesting andchanneling mechanism uses a hydrophobic field for fluid harvesting andhydrophilic channels for droplet movement onto the nano-sensor. Thismechanism makes the inventive system practical for creating a veryinexpensive, scalable manufacturable COVID-19 test that does not requireany blood or the administration of the test by a skilled technician,nurse or healthcare provider.

An embodiment uses a nano-scale fluidic biosensor technology with aunique moisture droplet harvesting and channeling structure. Thisstructure unlocks the use of the nano-scale sensor for detectionpossibly down to single molecules of target biomarkers. This enables thedetection of even very low concentrations of antibodies, proteins andother chemical biomarkers present in any body fluid without the drawingof blood.

A non-limiting embodiment builds on the sweat chemistry sensortechnology described in PCT/US19/45429, METHODS AND APPARATUS FOR AWEARABLE ELECTRONIC DIGITAL THERAPEUTIC DEVICE invented by Daniels andpublished Apr. 10, 2020, which is incorporated by reference herein inits entirety. The embodiment is a COVID-19 testing system that can bemass produced on readily available high volume manufacturing equipmentin the millions of units needed for mass population testing. Anembodiment of the testing system uses a nano-scale fluidic biosensorwith a unique moisture droplet harvesting and channeling structure. Thisstructure enables the use of the nano-scale sensor for detectingCOVID-19 biomarkers in a body fluid sample, such as breath condensate.This system enables the detection of analyte(s) of antibodies, proteins,RNA and other chemical COVID-19 biomarkers without the drawing of blood,expensive equipment or technically trained personnel. The proposedsystem can be configured as at least a first pass go/no-go test that candetermine who should be more accurately tested by the conventionaltesting methodologies.

COVID-19 is a new disease with uncertain knowledge about how it spreadsand the severity of the illness it causes. The virus is thought tospread mainly from person-to-person, between people who are in closecontact with one another. Respiratory droplets produced when an infectedperson coughs, sneezes or talks can land in the mouths or noses ofpeople who are nearby or possibly be inhaled into the lungs. COVID-19can be transferred by touching a surface contaminated with the virusthen touching one's eyes, nose or mouth. Recent studies suggest thatCOVID-19 may also be spread by people who are contagious butasymptomatic.

Mathematical models indicate that one-time lockdown interventions maynot be sufficient to keep the spread of COVID-19 from overwhelming theUS critical care capacity. Although seasonal variation in transmissionmay help slow the spread during the summer months, this could also leadto a dramatic increase in COVID-19 infections in the autumn. Researchersbelieve that intermittent distancing measures can maintain control ofthe epidemic, but without other interventions, these measures may benecessary into 2022 (see, Social distancing strategies for curbing theCOVID-19 epidemic, Kissler et al. pre-peer-reviewed online release,https://www.medrxiv.org/content/10.1101/2020.03.22.20041079v1.full.pdf).

Modeling studies and reports from the Wuhan outbreak indicate that evenin developed countries with good healthcare infrastructure, the capacityfor critical care response caused by COVID-19 infections can be exceededmany times over if distancing measures are not implemented quickly orstrongly enough¹. The modeling studies are showing that to keep criticalcare capacities from being overwhelmed, prolonged or intermittent socialdistancing may be necessary (see, Li R, Rivers C, Tan Q, Murray M, TonerE, Lipsitch M. 2020 The demand for inpatient and ICU beds for COVID-19in the US: lessons from Chinese cities).

So in order to slow the spread of the virus, governments have institutedstay at home policies, requiring billions of people around the world tostop their usual employment, entertainment and socializing activities.

This drastic shutdown of business has been a major element in combatingthe COVID-19 pandemic. However, the economic fallout is becominguntenable. As of April 16, there are more than 22 million people in theUS who have filed for unemployment. The financial markets are beingaffected around the world with unprecedented volatility, and governmentsare now trying to find the balance to combat the spread of the virusthrough country-wide lockdowns while allowing for some easing of thelockdowns to avoid possible economic collapse. A key question forgovernment leaders is: how do we safely restart our economies?

There is a growing consensus that testing for biomarkers that indicateexposure, infection and recovery from COVID-19 should be used to enablea safer and more efficient restart to regional, national and the globaleconomy, while minimizing the threat of an out-of-control spread of thevirus. The restarting of the global economy must be done safely toprevent avoidable suffering and an exacerbation of the socioeconomicdamage already caused by the pandemic. However, restarting must also bedone efficiently and as soon as practical to prevent a prolongedrecession and possible global depression that could take even more of asevere toll especially on poor and economically-at-risk people.

Testing for COVID-19 has become a very important tool in the arsenal forcombating the virus and enabling a safe restarting of the economy.Antibody testing, in particular, could determine who in the populationmay now be immune to reinfection or who is still susceptible to thevirus infection and a threat of infecting others. Importantly, knowingwho has adequate antibody protection against Covid-19 could more safelyenable the re-employment of a growing workforce of protected individualsand consumers while those who remain at risk of infection andtransmission can be kept sequestered until a vaccine or other mechanismis developed.

Tools for early diagnostics, infection prevention, monitoring andcontrol have become critical need items due the global outbreak of theCOVID-19 virus. In the past few months the world has learned a lot aboutthe virus that is causing the current pandemic, but still there is muchto learn about how to prevent exposures, identify who needs immediatetreatment, and how to slow the virus spread.

Most reasonable estimates for when a vaccine might be available forwidespread deployment range from 18 months to as much as two-threeyears. The need for mass testing for COVID-19 will last at least untilthere is a vaccine. Most likely even once this current pandemic isconquered the need to quickly spin up mass testing capabilities willalways be part of a country's long term strategic health plan.

Even after the first surge of the virus has passed, in order to properlydimension and locate resources for the next virus wave, governments andhealthcare providers need to determine who still needs to bequarantined, who currently has the active virus, and who has alreadyrecovered from the virus. Early detection mechanisms for all these casescan be used to significantly limit the virus spread and preventoverwhelming a state's or country's healthcare system.

There are several detection systems under development by companies andgovernment agencies around the globe. These systems detect severaldifferent biomarkers indicative of the COVID-19 virus.

For example, RayBiotech of Georgia offers lateral flow devices for thedetection of IgG and IgM antibodies to the coronavirus N-protein inserum, plasma, and peripheral blood. Such a lateral flow device issimilar to a home pregnancy test, but requires the drawing of bloodthrough a finger pin prick.

Other Covid-19 biomarker testing systems include a RT-PCR Nucleic Acid(Real Time) detection system that can be used for detecting anucleocapsid protein N gene from samples taken by human throat swabs andalveolar lavage.

Measurement of human IgG antibody can be performed for COVID-19 in serumor plasma using an ELISA testing methodology. The enzyme-linkedimmunosorbent assay (ELISA) typically uses a solid-phase enzymeimmunoassay to detect the presence of a target biomarker ligand orprotein.

There are several other detectable biomarkers that can help identifyexposure, incubation, infection, and post-recovery. Proteins, such as anucleocapsid protein (N-protein) that binds to the coronavirus RNAgenome creating a shell around the enclosed nucleic acid can be used asa detectable biomarker. Human ACE-2 (angiotensin-converting enzyme 2) isan integral membrane protein that serves as the initial attachment pointfor COVID-19 and can also be used as a detectable biomarker. To combatCOVID-19 and aid in drug discovery, protein profiling can be used tounderstand the host response to infection and identify potentialbiomarkers for drug development.

During the course of the progression of the virus in the human host,changes to the immune system can also be used as detectable biomarkers.For example, during infection of COVID-19, high levels of inflammatorycytokines include interferons, interleukins, chemokines,colony-stimulating factors, and tumor necrosis factors are oftendetectable and contribute to the symptoms of coronavirus infection.Recent research is showing that the overproduction of pro-inflammatorycytokines can result in a “cytokine storm,” where inflammation spreadsthroughout the body carried by the circulatory system. A commonconsequence of a cytokine storm is acute lung injury, which can thenresult in a more severe form called acute respiratory distress syndrome.Proteins and other biomarkers resulting from a cytokine storm can beused to help monitor and predict the progression of the virus.

An embodiment of the inventive COVID-19 testing system could be usefulfor creating a low cost, accurate, and easy-to-use testing system forsome or all of these COVID-19 indicating biomarkers. Such a system hasmultiple utilities including contact tracing, diagnosing, diseaseprogression monitoring and predictive machine learning population dataanalysis.

Blood tests and nasal swabs are now being used for testing for certainCOVID-19 biomarkers. The accuracy and sensitivity of these tests isstill lower than optimum, and typically require a technician or nursewith elaborate personal protection equipment to obtain the sample fromthe individual being tested. Then, the equipment that is used to performthe tests requires a skilled technician and is typically doneserially—one sample at a time. The inventive COVID-19 testing system isa low cost, mass producible technology platform that can be configuredto test for multiple COVID-19 biomarkers, and be deployed quickly forat-home testing, as well as for clinical and drug discovery use. Such amultiple biomarker testing system should have better statistical resultsthan the single biomarker tests currently being done as the standard ofcare.

There is the need for a low cost, accurate, easy-to-use testing systemfor COVID-19 that ideally can be mailed out and self-administered athome. For example, current testing protocols require a nasal swab forRNA testing to show active infection or a sample of blood be taken froma person in order to test for sufficient antibodies to the COVID-19virus for immunity. These tests typically require breaking sequestrationand traveling to a testing site where a technician, nurse or otherhealthcare provider administers the test. We propose a testing systemthat can be used as a first pass go/no go assessment to first see if amore elaborate and expensive testing methodology is warranted. Forexample, an inexpensive, easy-to-use testing system that can be done athome and finds a low concentration of COVID-19 antibodies present in thebreath or sweat can then be used as the impetus for the individual to goto a testing facility for a more accurate determination of the person'simmunity to further COVID-19 infection.

FIG. 5 is a side view of a wearable electronic breath chemistry sensor.FIG. 6 is a top view of a wearable electronic breath chemistry sensor.The biometric sensor is tuned to detect at least one biometric indicatorassociated with the presence of COVID-19 antigen, RNA and/or antibody. Adroplet collector draws EBC droplets into a transfer aperture. Thesensor is wet by the droplet and then the droplet is drawn throughwicking into wicking/evaporation materials. A continuous flow of freshdroplets passes over the sensor. A hydrophobic field encourages sweat tobead and migrate to hydrophilic channels. Tapered hydrophilic channelsuse surface tension to draw sweat into the sweat transfer aperture.Hydrophobic and hydrophilic screen printable inks are available fromcompanies such as Cytonix and Wacker.

FIG. 7 is an isolated view of an Exhaled Breath Condensate (EBC) dropletsample collector. FIG. 8 is a top view showing a step for forming theEBC droplet collector. FIG. 9 is a top view showing another step forforming the EBC droplet sample collector. FIG. 10 is a top view showingstill another step for forming the EBC droplet sample collector. FIG. 11is a top view showing yet another step for forming the EBC dropletsample collector. FIG. 12 illustrates the EBC sample collector showingEBS droplets. In accordance with a non limiting exemplary embodiment, anat-home, triage COVID-19 testing system uses Exhaled Breath Condensate(EBC) for an analyte fluid sample. Breath is an exceptional source ofvirus antigens, antibodies and RNA. EBC can be analyzed using establishmethods including Lateral Flow Assay, Nano-scale Bioreceptor andPhotonic Quantitative Assay. EBC produces much cleaner samples to testthan nasal swabs, is non-invasive and easier than drawing blood.However, collecting EBC usually requires a big, expensive chiller and isalways done in a clinical setting.

There is a great push throughout the world to develop adequate testingfor the COVID-19 virus. A conventional PCR test detects pieces of deadvirus from nasal swab or sputum. The test determines if a person isinfectious. The test is expensive, requires trained personnel andmachines and there is a delay in obtaining the test results due tocollection, transportation and processing of samples. PCR also requiresa lot of chemical and results in a lot of false negatives. Antibody testdetects the body's immune response to the virus. It requires a bloodsample. Antibody tests can be relatively fast and does necessarilyrequire trained personnel. False positives are frequent because otherviruses could be causing the antibodies.

EBC has been used for rapid detection of microbial DNA and RNA todemonstrate bacterial and viral lung infections. (see, Xu Z, Shen F, LiX, Wu Y Chen Q, Jie X, et al. Molecular and microscopic analysis ofbacteria and viruses in exhaled breath collected using a simpleimpaction and condensing method. PLoS One 2012; 7:e41137.https://www.ncbi.nlm.nih.gov/pubmed/22848436).

A nasal swab sample often contains a lot of background biologicalmaterials making it harder to identify the RNA of the virus because ofother molecules present in the sample. Breath condensate is naturallyenriched with viruses and confounding molecules are at much lowerconcentrations. (see,https://www.zimmerpeacocktech.com/products/electrochemical-sensors/covid-19-and-pcr-on-the-breath/).

Antibodies are present in breath vapor. IgA antibodies are found inareas of the body such the nose and breathing passages. IgG antibodiesare found in all body fluids and are the most common antibody (75% to80%), that are very important in fighting bacterial and viralinfections. IgE antibodies are found in the lungs, skin, and mucousmembranes. (see, https://www.uofmhealth.org/health-library/hw41342).

Virus Antigens are found in Airway Lining Fluid (ALF). EBC is anon-invasive method of sampling airway lining fluid (ALF). Constituentsof ALF are representative of the respiratory tree lining fluids. ALF isa measure of the concentration of biomarkers directly influenced byrespiratory cells. (see, Exhaled breath condensate: a comprehensiveupdate, Ahmadzai, et al., Clinical Chemistry and Laboratory Medicine(CCLM) 51, 7; 10.1515/cclm-2012-0593).

Although EBC could be an exceptional source of biomarkers indicating thestages of infection and recovery from the COVID-19 virus, as well asother medical and fitness uses, the conventional equipment for obtainingan EBC fluid sample is big and expensive, and is only used in clinicalsetting. The conventional equipment requires a chiller and is designedfor relatively large sample collection. This makes conventional EBCsampling equipment unsuited for at-home testing.

An embodiment of the inventive EBC sample collector includes ahydrophobic field that causes vapor to bead up into droplets.Hydrophilic channels coalesce and transfer the droplets to form anaccessible EBC fluid sample. The hydrophobic field and hydrophilicchannel can be screen printed or otherwise coated on a thermal massaluminum sheet substrate. This substrate can be chilled prior to usingthe testing system to improve EBC collection. The inventive EBC samplecollector makes the low cost Lateral Flow Assay and other testingsystems workable for at-home triage testing. The CDC says it isessential to quickly develop inexpensive screening test. The inventiveEBC sample collector makes such screening testing viable for massdeployment to large segments of the population. There is no need tobreak sequestration. No skilled technicians, clinics or lab equipmentare needed. Very high volume existing manufacturing methods can bemodified to product multiple-up (many at once) screen printed EBC samplecollectors. A low-cost aluminum substrate acts as thermal mass and canbe chilled for faster droplet harvesting. Batch fabrication can be usedto manufacture multiple-up LFA modules on a sheet with a format that isquickly adaptable to ultra-high-volume Roll-to-Roll manufacturing.

FIG. 13 illustrates the EBC sample collector applied to an LFA testingsystem. FIG. 14 is an exploded view showing the screen printedhydrophilic channels, screen printed hydrophobic field and thermal masssubstrate of the EBC sample collector. A first emitter/detector pair areused to determine if the novel coronavirus N protein at the test line(T) has been bound by the IgM-IgM complex. A second emitter/detectorpair are used to determine if free anti-human IgM antibody has beenbound to the anti-mouse antibody at the control line (C) confirming thatthe fluid sample has transversed through the transfer medium and thetest has been correctly performed.

In accordance with an embodiment, a method of forming a biomarkertesting system comprising forming an exhaled breath condensate fluidsample collector. Forming the exhaled breath condensate fluid samplecollector comprise the steps of providing a substrate, coating ahydrophobic field on the substrate, and coating at least one hydrophilicchannel on the substrate. The hydrophobic field is for receiving bodyfluid vapor and forming a fluid droplet from the received body fluidvapor and hydrophilic channel is for receiving the fluid droplet andchanneling the fluid droplet towards a testing system. At least onefluid sample draining hole may be formed at an end of the hydrophilicchannel for draining the fluid droplet through the at least one fluidsample draining hole onto a sample receiving structure of the testingsystem.

At least one photoemitter and one photodetector may be provided wherethe photoemitter emits radiation towards the biomarker testing zone andthe photodetector receives radiation from the biomarker testing zone.FIG. 15 is an exploded view showing the constituent elements of a LFA.FIG. 16 illustrates an embodiment of the LFA including photonicemitter/detector electronics. In immunochromagtography, a captureantibody is disposed onto a surface of a porous membrane, and a samplepasses along the membrane. Analyte in the sample is bound by theantibody which is then coupled to a detector reagent. As the samplepasses through the area where the capture reagent is disposed, theanalyte detector reagent complex is trapped, and a color develops thatis proportional to the analyte present in the sample. The photonicsemitter/detector pair enable the proportional quantitative measurementof the analyte where the analyte concentration if the fluid sample isdetermined from an intensity or counting of received photons at thedetector.

The solid-phase lateral-flow test platform is an example ofimmunochromatography that is widely used for home pregnancy testing.Lateral flow tests have benefited from the use of sol particles aslabels. The use of inorganic (metal) colloidal particles are typicallyused as a label for immunoassays and several techniques are used tomeasure the amount of bound conjugate. These include naked eye,colorimetry and atomic absorption spectrophotometry. Colorimetry appliesthe Beer-Lambert law, which states that the concentration of a solute isproportional to the absorbance. At higher antigen concentrations, theresults of immunochromagtography can be read by the naked eye (e.g., thetypical home pregnancy test). For lower concentrations, colorimetry hasbeen shown to be more than 30 times more sensitive than reading by thenaked eye.

In accordance with an embodiment, immunochromatography is used to detectthe present of a COVID-19 analyte. Generally, immunochromatography isthe separation of components in a mixture through a medium usingcapillary force and the specific and rapid binding of an antibody to itsantigen. A dry transfer medium is coated separately with novelcoronavirus N protein (“T” test line) and anti-mouse antibody (“C”control line). Free colloidal gold-labeled anti-human IgM are in arelease pad section (S). The inventive vapor coalescence and dropletharvesting structure are used to obtain a fluid sample of breathcondensate. This fluid sample is applied to the release pad section. Theanti-human IgM antibody the binds to at least some of the IgM antibodies(if any are present), forming an IgM-IgM complex. The fluid sample andantibodies move through the transfer medium via capillary action. Ifcoronavirus IgM antibody is present in the fluid sample, the novelcoronavirus N protein at the test line (T) will be bound by the IgM-IgMcomplex and develop color. If there is no coronavirus IgM antibody inthe sample, free anti-human IgM does not bind to the test line (T) andno color will develop. The free anti-human IgM antibody will bind to theanti-mouse antibody at the control line (C) so that the control linedevelops color confirming that the fluid sample has transversed throughthe transfer medium and the test has been correctly performed.

FIG. 17 illustrates the EBC sample collector applied to a nanoscalebioreceptor testing system and showing a pull tab for holding backcollected droplets on the sample pad. FIG. 18 is a perspective viewshowing the EBC sample collector applied to a testing system. FIG. 19 isan isolated view showing the pull table disposed between the sample padand conjugate release pad. The EBC fluid sample will be collected oversome time period, collected from the hydrophobic field through thehydrophilic channels, and flowing through the fluid sample drain holesto build up in the sample pad. A typical LFA uses about 3 drops ofbuffered sample that is added to the sample pad. In accordance with anexemplary embodiment, buffer materials and surfactants can beincorporated in dry form into the sample pad so that the EBC sample,which is mostly water, will be suitable as a directly applied fluidsample without requiring the addition of a fluid buffer. When a personis at rest, there is about 17.5 ml of EBC produced per hour (see, Howmuch water is lost during breathing?, Zielinski et al., PneumonolAlergol Pol 2012; 80(4):339-342). There are 20 drops per milliliter. So,every hour at rest there is the potential to collect about 350 drops ofEBC. A collection efficiency by the EBC sample collector of only about3% should provide an adequate number of EBC droplets for a fluid samplein about 15 minutes from an individual at rest.

In accordance with an embodiment, an apparatus for detecting a biomarkercomprises a droplet harvesting structure for converting breath vapor toa fluid droplet for forming a fluid sample and a testing system having abiomarker testing zone for receiving the fluid sample and detecting abiomarker. The droplet harvesting structure may include at least one ofa hydrophobic field for receiving the breath vapor and forming the fluiddroplet from the received breath vapor and hydrophilic channels forreceiving the fluid droplet and channeling the fluid droplet towards thetesting system. A fluid dam member may be provided disposed between thedroplet harvesting structure and the biomarker testing zone.

The testing system may comprise a fluidic lateral flow assay including asample pad for receiving the fluid sample potentially containing abiomarker analyte, a conjugate release pad, a flow membrane and anadsorbent pad for receiving and flowing the fluid sample to detect thepotential biomarker analyte from the sample source. A fluid dam memberdisposed between the sample pad and the conjugate release pad, the fluiddam including a pull tab structure to enable a user to remove the fluiddam member and allow the flow of the fluid sample from the sample pad tothe conjugate release pad.

In order to flush the EBC fluid sample through the test system, thefluid dam is provided to hold back the EBC on the sample pad as itaccumulates. The fluid dam may be, for example, a piece of siliconecoated release paper forming a pull tab that is disposed between thesample pad and the conjugate release pad. The pull tab is held in placeon the top adhesive and allows for the accumulation of the EBC fluidsample. After an adequate amount of time has passed to saturate thesample pad with enough EBC fluid sample, the user pulls the pull tab outcausing the EBC fluid sample to flow from the sample pad to theconjugate release pad. This enables the EBC fluid sample to flushthrough the various constituents of the testing system by capillaryaction. The EBC fluid sample is allowed to build up on the sample pad sothat removing the pull tab releases sample flow all at once to ensuresadequate sample flow and promotes testing consistency.

FIG. 20 is an isolated view of a screen printed droplet sample collectorwith a fluid transfer aperture. FIG. 21 is a cross-section view showinga fluid sample collected from the EBC sample collector flowing between aphotonics emitter/detector pair. In this embodiment, another body fluid,such as sweat, might be used with the EBC sample collector configured toharvest sweat droplets from the skin instead of coalescing breath vaporinto exhaled breath condensate. It is noted that any of the embodimentsand innovations described herein may be useful for other medical andfitness uses, for other disease or virus testing or biometric detectionin addition to or instead of the described use for COVID-19 testing.

The manufacturing techniques, equipment and materials for mostcomponents of an embodiment of the inventive COVID-19 testing system arereadily available and very well known. For example, to create our fluidharvesting and droplet channeling structure, screen printing is used topattern hydroscopic and hydrophobic inks sourced from a company such asCytonix and Wacker. There is no shortage of manufacturing capacityneeded to quickly screen print for the hundreds of millions of testingunits needed. The fluidic biosensor component can be manufactured usinghigh throughput equipment available from a company such as Nano-Ops,Boston, Mass., and the chemistry for functionalizing the biosensor canbe obtained from a company such as RayBiotech, Peach Tree Corners, Ga.Other necessary manufacturing steps, such as wire bonding and printedcircuit board fabrication will make use of the same ubiquitous machinesthat are similarly purposed for semi-conductor and circuit boardelectronics.

FIG. 22 shows the side views of the steps for building up an LFA testingsystem. FIG. 23 shows the top view of the steps for building up an LFAtesting system. FIG. 24 shows a 4×9 ganged multiple-up sheet of LFAtesting systems formed as a batch. FIG. 25 shows a roll-to-rollmanufacturing process for forming a roll of bottom adhesive/backingsubstrate/top adhesive.

FIG. 26 is a perspective view illustrating the bottom adhesive/backingsubstrate/top adhesive stack. FIG. 27 shows a roll-to-roll manufacturingprocess for forming the constituent elements of an LFA on a roll ofbottom adhesive/backing substrate/top adhesive. FIG. 28 shows the LFAtesting system formed by the roll-to-roll process cut from a continuousroll and showing a section of top adhesive for adhering the LFA testingsystem to a separately formed ENC sample collector. FIG. 29 shows theLFA testing system formed by the roll-to-roll process cut from acontinuous roll and showing a section of bottom adhesive for stickingonto a wearable garment such as a face mask. FIG. 30 shows a sheet ofsubstrate with a hydrophobic field coating on a thermal mass substratewith droplet collection holes. FIG. 31 shows the sheet of substrate withthe hydrophobic field coating on the thermal mass substrate with dropletcollection holes having a coating of hydrophilic channels.

FIG. 32 shows the EBC sample collector and testing system withelectronics for wireless data acquisition and transmission along withseparate trusted receiver and public blockchain data path and storage.Villanova University recently published an example of utilizingblockchain to help medical facilities track coronavirus cases globally.A private blockchain is shared among medical facilities around the worldto publish coronavirus test results between doctors on a trusted,immutable ledger. IoT and AI are used to survey public spaces wherehigh-risk gatherings can take place and trigger alerts over theblockchain. (see,https://www.villanova.edu/university/experts/spotlight-detail.html?spotlight=7180).

In accordance with an exemplary embodiment, the EBC collecting systemwith biomarker detection can utilize self-reporting or automatic datacollection to be usable with a new or existing APPs for contact tracingand electrical medical records. The acquired data can anonymized andencrypted at the source (e.g., on the electronics associated with thetesting system). A first data stream/data base allows a trusted receiverto access patient identifying data while a second data stream/data baseprovides anonymized data that can be provided as open source or otherdata transmission, storage and utilization mechanisms withoutidentifying who the source of the data is from.

The inventive testing system has the potential to be very low cost,shippable in a conventional envelop for mass distribution to everyhousehold in a target region, state or country. This enables a muchhigher percentage of the population to undergo at least the baselinetesting indicating if they should follow up with a visit to a drivethrough, hospital or clinic testing facility for more elaborate testing.

The inventive COVID-19 testing system can have the capability of testingtwo or more of the virus biomarkers. For example, RNA testing can becombined with antibody testing. By testing for these two biomarkers thepotential for false negatives is significantly statistically reduced andlikely will be a more preferable methodology.

The proposed COVID-19 testing system can be incorporated into personalprotection equipment, such as masks, or provided as a patch that isstuck onto the body, or provided as a stand-alone test unit, similar toa home pregnancy test.

The testing system can include wireless communications capabilities,such as RFID and Bluetooth. This will enable, for example, test data tobe used along with GPS location information to assist in contact tracingand further quicken the ability of a growing segment of the populationto return to work and restart economic activities, and to also determinethrough real-time contact tracing who might have been exposed to thevirus.

As an enhancement to the basic system, biometric data can be acquiredand used for the public good. The collection of biometric informationcarries with it the burden of privacy issues. There can be consideredtwo uses for a patient's biometric data: Patient monitoring forprevention and treatment; and Population studies to improve globalhealthcare. The inventive system uses separately created and maintaineddata bases.

The biometric parameters such as those described herein with regards tothe embodiments can also be detected, logged and/or transmitted,enabling a detailed history of the patient's disease progression,therapy, course of treatment, measured results of treatment, etc., andcan be made available to improve the care given to the particularpatient, and in the aggregate, provide significant data along with thatof other patients, to assist in new drug discovery, treatmentmodifications, and a number of other advantages of the beneficial cyclecreated by detection, transmission, storage and analysis of biometricdata taken directly from the patient during the course of drug therapyand/or other treatments.

FIG. 33 shows the manufacturing processes for a heat bonded face mask.FIG. 34 shows the fabric, filter and other layers bonded through aroll-to-roll lamination process ore individually cut into blanks forforming a pre-form mask stack. FIG. 35 shows other materials such asbiological reactive silver fabric and hot melt adhesive of the pre-formmask stack. The highly contagious and deadly effects of COVID-19 haveresulting in an increased need for personal protective masks. Disposablemasks are a good solution for healthcare providers, police and otherswho's job put them into constant contact with individuals who may or maynot have the virus. The ability to change out a disposable mask betweenpatients, for example, ensures that a doctor or nurse will have a fresh,clean, uncontaminated mask to better protect themselves and protecttheir patients from the spread of the virus. However, disposable masksare not a good solution for the general population. The cost and wasteassociated with a disposable mask makes it a poor solution for mostpeople. Rather, what is needed is a mask that is low cost, easy tomanufacture and ideally can be sterilized in a conventional home clotheswashing machine and dryer.

FIG. 36 is an exploded view of a mask stack. FIG. 37 shows the foldlines of the mask stack for first and second heat press operations. FIG.38 shows the folded, pressed and heat bonded mask. FIG. 39 shows theattachment of the EBC collector and testing system to the folded mask.FIG. 40 shows the step of turning the folded mask inside out to disposethe EBC collector and testing system on the inside of the mask. FIG. 41shows a heat press operation to bond elastic straps onto the foldedmask. FIG. 42 shows the mask with the EBC collector and testing systemdisposed inside the mask within the concentrated atmosphere of exhaledbreath. FIG. 43 shows a conventional bendable metal nose seal that isdisposed within the folds of the mask at a location corresponding to thebridge of a user's nose. FIG. 44 shows a replaceable adhesive nose stripthat is disposed on the outside of the folds of the mask at a locationcorresponding to the bridge of a user's nose, and FIG. 45 shows thecomponents of a magnetic removable nose seal.

FIG. 46 is an exploded view of a testing system including a dissolvableflow dam that holds back collected EBC on the sample pad until enoughhas been accumulated to be released onto the conjugate release pad andflush the fluid sample through the components of the testing system. Afluid dam member may be disposed between the droplet harvestingstructure and the biomarker testing zone, wherein the fluid dam memberincludes at least one of a removable moisture resistant sheet member anda dissolvable film for accumulating the fluid sample from the dropletharvesting structure and releasing the accumulated fluid sample to flowto the biomarker testing zone. The fluid sample testing system comprisesa fluidic lateral flow assay including a sample pad for receiving thefluid sample potentially containing a biomarker analyte as the secondbiomarker, a conjugate release pad, a flow membrane and an adsorbent padfor receiving and flowing the fluid sample to detect the potentialbiomarker analyte from the sample source. The fluid dam member may bedisposed between the sample pad and the conjugate release pad, the fluiddam including a pull tab structure to enable a user to remove the fluiddam member and allow the flow of the fluid sample from the sample pad tothe conjugate release pad. At least one photoemitter and onephotodetector may be provided, wherein the photoemitter emits radiationtowards the biomarker testing zone and the photodetector receivesradiation from the biomarker testing zone.

FIG. 47 is an isolated view showing the dissolvable flow dam insertedbetween the sample pad and the conjugate release pad. FIG. 48 is anisolated view showing after the dissolvable flow dam has been dissolvedaway to release the accumulated fluid sample from the sample pad to theconjugate release pad. The inventive at-home testing system can be usedfor COVID-19, other virus, bacterial, environment, cancer, asthma,diabetes, fitness, or other medical use-case. The basic premise is tocollect Exhaled Breath Condensate (EBC) and Exhaled Breath Aerosols(EBA) using a face mask. The EBC is collected through ahydrophobic/hydrophilic droplet harvesting structure and channeled ontoa testing system (e.g., Lateral Flow Assay or Electronic Biosensor). Toeffectively collect and accumulate EBC, a dissolvable material may beused for regulating capillary fill time. This allows holding back theflow of the liquid sample (EBC) from the droplet harvesting structureuntil enough sample is accumulated to flush the liquid via capillaryaction through the test system. For capturing EBA, suspending dropletsand aerosol particulate on the surface or into a film of a dissolvablefilm can be used where the surface of the film is tacky so that exhaledparticulate during breathing or coughing will stick to the adhesivesurface. If the film is also water soluble, breath droplet will also beadsorbed into the film. This COVID-19 testing system can be deployed forusing EBC for screening (that is, a go/no-go triage test) and if the EBCtest indicates a positive detection of a target biomarker (e.g.,COVID-19 antibody or RNA), then the mask is shipped to a testing labwhere the captured EBA is analyzed FIG. 49 is an isolated view showing adissolvable EBC droplet and EBA particulate collector. FIG. 50 is across section side view showing a section of the dissolvable droplet andparticulate collector having particulate and droplets impinged on thesurface. In an enhanced version of the proposed testing system anaerosol particulate collection system is provided to capture virusbiomarkers exhaled or coughed by the test subject. The surfaces in allparts of the lung down to the alveoli are coated with an aqueous mucouslayer that can be aerosolized and carry along a variety of non-volatileconstituents. EBC and EBA are different types of breath matrices used toassess human health and disease state. EBA represents a fraction oftotal EBC, and is targeted to larger molecules, such as fatty acids andcytokines, as well as cellular fractions, proteins, viruses, andbacteria instead of the gas-phase (see, Wallace M A G, Pleil J D.Evolution of clinical and environmental health applications of exhaledbreath research: Review of methods and instrumentation for gas-phase,condensate, and aerosols. Anal Chim Acta. 2018; 1024:18-38.doi:10.1016/j.aca.2018.01.069).

FIG. 51 is a cross section side view showing the section of thedissolvable droplet and particulate collector having particulateembedded into the dissolvable capture film and droplets dissolved intoand causing a detection reaction with a detection reagent of thedissolvable capture film. FIG. 52 is a top view showing the inventivetesting system including a dissolvable EBC droplet and EBA particulatecollector having captured aerosol droplets and aerosol particulate. Theparticulate capture mechanism can be dissolvable film that has a stickysurface and may include a visual detection reaction to one or moretarget biomarkers. A soluble biomarker that reacts with the visualdetection chemical generates a visual indication of the biomarkerpresences in the EBA. A non-soluble particulate is captured on thesticky surface and becomes embedded into the capture film so it can beeasily shipped to a lab for analysis. As an example, if the EBC testingsystem is used for at-home screening, a positive test result for the EBCtarget biomarker can be used to prompt the test subject to mail back thetesting system so that the captured particulate from the EBA sample canbe further analyzed with more sophisticated laboratory equipment.

The inventive system for detecting a biological agent from the breath ofa test subject comprises an exhaled breath condensate droplet harvesterfor coalescing breath vapor into droplets to form a fluid biologicalsample, a testing system for receiving the fluid biological sample fromthe breath droplet harvester and testing for a target analyte, and awireless communication electronic circuit for detecting a result of thetesting for the target analyte and communicating the result to awireless receiver.

An exhaled breath aerosol capture system can be provided comprising asheet member having a surface for receiving exhaled breath aerosolcomprising at least one of a particulate and a droplet. The surface canbe non-soluble, pressure sensitive adhesive or an exposed portion of adissolvable film formed on, coated, adhered to or integral with thesheet member. The dissolvable film has a composition effective forreceiving and capturing the at least one of a particulate and a dropletby at least one of embedding or dissolving the at least one of aparticulate and a droplet onto the surface or into the dissolvable film.

At least one of the surface and the dissolvable film includes a reagentfor reacting with the at least one particulate and droplet for detectingfor the presence of a target analyte in the at least one particulate anddroplet.

FIG. 53 is an isolated perspective view showing the dissolvable EBCdroplet and EBA particulate collector having captured aerosol dropletsand aerosol particulate. FIG. 54 is a top view showing the inventivetesting system including a dissolvable EBC droplet and EBA particulatecollector before capturing aerosol droplets and aerosol particulate.FIG. 55 is a top view showing the inventive testing system including adissolvable EBC droplet and EBA particulate collector after capturingaerosol droplets and aerosol particulate.

In a further enhanced version of the proposed COVID-19 test system, anano sensor array can be included along with the EBC and/or EBAcollection systems to also test for VOCs, nitric oxide and other gaseousbiomarkers specific to virus and/or accompanying changes in the body inresponse to exposure to COVID-19. FIG. 56 is a top view showing theinventive testing system including a dissolvable EBC droplet and EBAparticulate collector installed onto a face mask substrate along with aplurality of gas sensors for detecting volatile and gas constituents ofthe exhaled breath and/or ambient atmosphere. A common feature of theinflammatory response in patients who have actually contracted influenzais the generation of a number of volatile products of the alveolar andairway epithelium. These products include a number of volatile organiccompounds (VOCs) and nitric oxide (NO). These may be used as biomarkersto detect the disease. A research team has shown that a portable3-sensor array microsystem-based tool can detect flu infectionbiomarkers (see, for example, Gouma P I, Wang L, Simon S R, StanacevicM. Novel Isoprene Sensor for a Flu Virus Breath Monitor. Sensors(Basel). 2017; 17(1):199. Published 2017 Jan. 20.doi:10.3390/s17010199). The gas sensors can be connected with the sameelectronics and wireless communication system used by the otherbiometric detecting capabilities of the inventive testing system.

FIG. 57 is a cross section side view showing a section of thedissolvable droplet and particulate collector having particulate anddroplets impinged on the surface placed in a beaker of dissolvingliquid. FIG. 58 is a cross section side view showing a section of thedissolvable droplet and particulate collector having the particulatereleased into and the droplets dissolved into the beaker of dissolvingliquid. In a proposed use-case, the inventive testing system can bedistributed on a massive scale through the mail or courier systems of acountry, state or region. The inventive test system can be incorporatedinto a mask as shown or provided as a stand alone system that can beeasily retrofit into an existing mask. As an alternative to the EBCDroplet Harvester, and alternative mechanism can be used to collect theEBC. For example, in a hospital setting, EBC can be collected from theface mask used to administer oxygen or other gas to a patient. At home,EBC can be collected by exhaling into a chiller tube (not shown) orother breath vapor condensing system.

The dissolvable droplet and particulate collector can be mailed to atesting laboratory where it is analyzed for captured biomarkers.Particulate and/or droplets can be expelled by the test subject througha forced cough, deep airway exhalation, sneeze, or other respiratorymaneuver. In a triage or screening procedure, a large number of testingsystems can be distributed to a whole population or statisticallymeaningful sample of the population. If the EBC testing system indicatesa likelihood of COVID-19 current or prior infection (or other biologicalcondition), then the entire testing system kit or just the dissolvabledroplet and particulate collector can be sent to laboratory for morestringent analysis.

The dissolving liquid used by the laboratory (or other testing facility)for testing for target biomarkers may include reagents that changecolor, cause precipitation, amplification or otherwise assist in theidentification of the target biomarker captured by the dissolvabledroplet and particulate collector.

In accordance with a non-limiting exemplary embodiment, a system isprovided for detecting a biological agent from the breath of a testsubject comprises an exhaled breath condensate droplet harvester forcoalescing breath vapor into droplets to form a fluid biological sample,a testing system for receiving the fluid biological sample from thebreath droplet harvester and testing for a target analyte, and awireless communication electronic circuit for detecting a result of thetesting for the target analyte and communicating the result to awireless receiver. An exhaled breath aerosol capture system can beprovided comprising a sheet member having a surface for receivingexhaled breath aerosol comprising at least one of a particulate and adroplet. The surface can be non-soluble, pressure sensitive adhesive oran exposed portion of a dissolvable film formed on, coated, adhered toor integral with the sheet member. The dissolvable film has acomposition effective for receiving and capturing the at least one of aparticulate and a droplet by at least one of embedding or dissolving theat least one of a particulate and a droplet onto the surface or into thedissolvable film. At least one of the surface and the dissolvable filmincludes a reagent for reacting with the at least one particulate anddroplet for detecting for the presence of a target analyte in the atleast one particulate and droplet.

Turning to FIG. 59, this figure shows a block diagram of one possibleand non-limiting exemplary system in which the exemplary embodiments maybe practiced. In FIG. 59, a COVID-19 testing system (C19TS) 110 is inwireless communication with a wireless network 100. A C19TS is awireless COVID-19 testing system that can access a wireless network. TheC19TS 110 includes one or more processors 120, one or more memories 125,and one or more transceivers 130 interconnected through one or morebuses 127. Each of the one or more transceivers 130 includes a receiver,Rx, 132 and a transmitter, Tx, 133. The one or more buses 127 may beaddress, data, or control buses, and may include any interconnectionmechanism, such as a series of lines on a motherboard or integratedcircuit, or other optical communication equipment, and the like. The oneor more transceivers 130 are connected to one or more antennas 128. Theone or more memories 125 include computer program code 123. The C19TS110 includes a Target Biomarker Collection and Analysis (TBCA) module140, comprising the inventive COVID-19 testing system described herein.An embodiment of the TBCA also includes wireless communicationcapabilities comprising one of or both parts 140-1 and/or 140-2, whichmay be implemented in a number of ways. The TBCA module 140 may beimplemented in hardware as TBCA module 140-1, such as being implementedas part of the one or more processors 120. The TBCA module 140-1 may beimplemented also as an integrated circuit or through other hardware suchas a programmable gate array. In another example, the TBCA module 140may be implemented as TBCA module 140-2, which is implemented ascomputer program code 123 and is executed by the one or more processors120. For instance, the one or more memories 125 and the computer programcode 123 may be configured to, with the one or more processors 120,cause the COVID-19 testing system 110 to perform one or more of theoperations as described herein. The C19TS 110 communicates with Node 170via a wireless link 111.

The Node 170 is a base station (e.g., 5G, 4G, LTE, long term evolutionor any other cellular, internet and/or wireless network communicationsystem) that provides access by wireless devices such as the C19TS 110to the wireless network 100. The Node 170 includes one or moreprocessors 152, one or more memories 155, one or more network interfaces(N/W I/F(s)) 161, and one or more transceivers 160 interconnectedthrough one or more buses 157. Each of the one or more transceivers 160includes a receiver, Rx, 162 and a transmitter Tx, 163. The one or moretransceivers 160 are connected to one or more antennas 158. The one ormore memories 155 include computer program code 153. The Node 170includes a Data Acquisition and Storage (DAS) module 150, comprising oneof or both parts 150-1 and/or 150-2, which may be implemented in anumber of ways. The DAS module 150 may be implemented in hardware as DASmodule 150-1, such as being implemented as part of the one or moreprocessors 152. The DAS module 150-1 may be implemented also as anintegrated circuit or through other hardware such as a programmable gatearray. In another example, the DAS module 150 may be implemented as DASmodule 150-2, which is implemented as computer program code 153 and isexecuted by the one or more processors 152. For instance, the one ormore memories 155 and the computer program code 153 are configured to,with the one or more processors 152, cause the Node 170 to perform oneor more of the operations as described herein. The one or more networkinterfaces 161 communicate over a network such as via the links 176 and131. Two or more Nodes 170 communicate using, e.g., link 176. The link176 may be wired or wireless or both and may implement, e.g., an X2interface.

The one or more buses 157 may be address, data, or control buses, andmay include any interconnection mechanism, such as a series of lines ona motherboard or integrated circuit, fiber optics or other opticalcommunication equipment, wireless channels, and the like. For example,the one or more transceivers 160 may be implemented as a remote radiohead (RRH) 195, with the other elements of the Node 170 being physicallyin a different location from the RRH, and the one or more buses 157could be implemented in part as fiber optic cable to connect the otherelements of the Node 170 to the RRH 195.

The wireless network 100 may include a network control element (NCE) 190that may include MME (Mobility Management Entity)/SGW (Serving Gateway)functionality, and which provides connectivity with a further network,such as a telephone network and/or a data communications network (e.g.,the Internet). The Node 170 is coupled via a link 131 to the NCE 190.The link 131 may be implemented as, e.g., an S1 interface. The NCE 190includes one or more processors 175, one or more memories 171, and oneor more network interfaces (N/W I/F(s)) 180, interconnected through oneor more buses 185. The one or more memories 171 include computer programcode 173. The one or more memories 171 and the computer program code 173are configured to, with the one or more processors 175, cause the NCE190 to perform one or more operations.

The wireless network 100 may implement network virtualization, which isthe process of combining hardware and software network resources andnetwork functionality into a single, software-based administrativeentity, a virtual network. Network virtualization involves platformvirtualization, often combined with resource virtualization. Networkvirtualization is categorized as either external, combining manynetworks, or parts of networks, into a virtual unit, or internal,providing network-like functionality to software containers on a singlesystem. Note that the virtualized entities that result from the networkvirtualization are still implemented, at some level, using hardware suchas processors 152 or 175 and memories 155 and 171, and also suchvirtualized entities create technical effects.

The computer readable memories 125, 155, and 171 may be of any typesuitable to the local technical environment and may be implemented usingany suitable data storage technology, such as semiconductor based memorydevices, flash memory, magnetic memory devices and systems, opticalmemory devices and systems, fixed memory and removable memory. Thecomputer readable memories 125, 155, and 171 may be means for performingstorage functions. The processors 120, 152, and 175 may be of any typesuitable to the local technical environment, and may include one or moreof general purpose computers, special purpose computers,microprocessors, digital signal processors (DSPs) and processors basedon a multi-core processor architecture, as non-limiting examples. Theprocessors 120, 152, and 175 may be means for performing functions, suchas controlling the C19TS 110, Node 170, and other functions as describedherein.

In general, the various embodiments of the COVID-19 testing system 110can include, but are not limited to, wireless communication componentsused for Bluetooth, cellular telephones such as smart phones, tablets,personal digital assistants (PDAs) having wireless communicationcapabilities, portable computers having wireless communicationcapabilities, image capture devices such as digital cameras havingwireless communication capabilities, gaming devices having wirelesscommunication capabilities, music storage and playback appliances havingwireless communication capabilities, Internet appliances permittingwireless Internet access and browsing, tablets with wirelesscommunication capabilities, as well as portable units or terminals thatincorporate combinations of such functions.

FIG. 60 is a logic flow diagram for Applied Probabilistic Analysis toDetermine COVID-19 Exposure. This figure further illustrates theoperation of an exemplary method, a result of execution of computerprogram instructions embodied on a computer readable memory, functionsperformed by logic implemented in hardware, and/or interconnected meansfor performing functions in accordance with exemplary embodiments. Forinstance, the TBCA module 140 may include multiples ones of circuitelements for implementing the functions shown in the blocks in FIG. 59,where each included block is an interconnected means for performing thefunction in the block. At least some of the blocks in FIG. 59 areassumed to be performed by the C19TS 110, e.g., under control of theTBCA module 140 at least in part.

For the applied probabilistic analysis to determine COVID-19 exposure,Biomarker1 is tested (step one), Biomarker1 is tested (step four), andBiomarkerN is tested (step three) where N can be any number of multiplebiomarkers tested using the inventive testing system. If no targetbiomarker is detected (step three) then a Negative Test report isgenerated (step four). If any target biomarker is detected (step three)then probabilistic analysis may be performed depending simply on thedetected presence (yes/no) or quantitative analysis (e.g.,concentration) of the one or more detected biomarkers (step five). Ifthe probabilistic analysis does not exceed a threshold (step six) (e.g.,low concentration of a particular target biomarker, or the presence ofjust one weak biomarker indicating likely infection), then a Maybe Testreport is generated (step seven). If the probabilistic analysis doesexceed a threshold (step six) (e.g., high concentration of a particulartarget biomarker, or the presence of two or more biomarkers indicatinglikely infection), then a Positive Test report is generated (stepeight). The Test Report is then transmitted (step nine) (e.g., in amanner described herein or other suitable transmission mechanismincluding verbal, digital, written or other communication transmission).

The logic flow of FIG. 60 is implemented by a non-limiting embodiment ofan apparatus, comprises: at least one processor; and at least one memoryincluding computer program code, the at least one memory and thecomputer program code configured to, with the at least one processor,cause the apparatus to perform at least the following: detecting one ormore biometric parameters using a droplet harvesting structure forconverting breath vapor to a fluid droplet for forming a fluid sampleand a testing system having a biomarker testing zone for receiving thefluid sample and detecting the biometric parameter, where the biometricparameters are biomarkers dependent on at least one physiological changeto a patient in response to a concerning condition such as a virusinfection; receiving the one or more biometric parameters and applyingprobabilistic analysis to determine if at least one physiological changethreshold has been exceeded dependent on the probabilistic analysis ofthe one ore more biometric parameters; and activating an actiondepending on the determined exceeded said at least one physiologicalchange.

In accordance with an embodiment, a digital testing device is providedcomprising a biomarker testing device having one or more biometricdetectors each for detecting biomarkers as one or more biometricparameters. The biometric parameters are dependent on at least onephysiological change to a patient or test subject, such as theproduction of immune response chemicals, the presence in the body of anactive or deactivated virus or virus component, antibodies, antigens,virus RNA or DNA, or other biomarker inducing change. A microprocessorreceives the one or more biometric parameters and determines if at leastone physiological change threshold has been exceeded dependent on theone or more biometric parameters. An activation circuit activates anaction depending on the determined physiological change. The actionincludes at least one of transmitting an alert, modifying a therapeutictreatment, and transmitting data dependent on at least one physiologicalchange, the one or more biometric parameters, and the therapeutictreatment.

The at least one physiological change can also be in response to anapplied therapeutic treatment that is at least one of a pharmaceuticaltreatment and an electroceutical treatment that causes a change in thecondition of the patient enabling the monitoring of the body's responseto the applied therapeutic. The action can include transmitting analert, modifying a therapeutic treatment, and transmitting datadependent on at least one of the at least one physiological change, theone or more biometric parameters, and the therapeutic treatment. Themicroprocessor can analyze the one or more biometric parameters usingprobabilistic analysis comprising determining from a data set of the oneor more biometric parameters whether the data set is acceptable fordeciding that the at least one physiological change threshold has beenexceeded. The probabilistic analysis can further comprise applying astatistical weighting to each of the one or more biometric parameters,where the statistical weighting is dependent on a predetermined value ofa ranking of importance in detecting each of the at least onephysiological change for said each of the one or more biometricparameters relative to others of the one or more biometric parameters.

FIG. 61 is a logic flow diagram for Data Acquisition and Transmissionfor Trusted Receiver and Contract Tracing Uses. This figure furtherillustrates the operation of an exemplary method, a result of executionof computer program instructions embodied on a computer readable memory,functions performed by logic implemented in hardware, and/orinterconnected means for performing functions in accordance withexemplary embodiments. The performance of the Data Acquisition andTransmission for Trusted Receiver and Contract Tracing Uses flow can bedone at the testing system, Node, Smartphone, or combination ofcomponents located or associated with the test subject through the enduser(s) or final storage location(s) of the acquired data. The acquireddata can include patient or subject identifying information ranging fromname, GPS location, list of known contacts, prior medical history,demographics, etc. The Data Acquisition and Transmission for TrustedReceiver and Contract Tracing Uses can be done at a secure serverlocated anywhere on the network. For instance, the DAS module 150 mayinclude multiples ones of circuit elements for implementing thefunctions shown in the blocks in FIG. 59, where each included block isan interconnected means for performing the function in the block. Atleast some of the blocks in FIG. 59 are assumed to be performed by abase station such as Node 170, e.g., under control of the DAS module 150at least in part.

The digital testing system architecture, manufacturing methods, andapplications, can be used for capturing biometric data from the exhaledbreath of a test subject or patient. Biometric data can be captured andtransmitted continuously or at selected times with data access provideddirectly to a care-provider, enabling early diagnosis and ongoingmonitoring, and to a researcher to gain valuable insights and assistancethrough AI analysis. This data detection is direct from the exhaledbreath and can be provided through a wireless connection for Blockchainand AI database collection, access and analysis. The inventive digitaltesting system for biometric capture is adapted to mass production as aroll-to-roll manufactured testing device with embedded sensors andtransducers.

The Test Report is received (step one) (e.g., from a Smartphonetransmission from the patient or test subject). If the report isintended to be sent to a trusted receiver (step two), such as apatient's healthcare provider or insurance company, then an encryptedreport can be generated (step three) and transmitted to the trustedreceiver that includes patient identifying information. If the report isnot for a trusted receiver (step two) but instead is to be used forcontact tracing (step four), then only the data required for ContactTracing is transmitted to a Contact Tracing APP (step five). The ContactTracing APP may be, for example, a system provided for identifying andnotifying people who have come in contact with the test subject orpatient within a given time prior or since testing positive or may befor one or more target biomarkers. If the report is not for a trustedreceiver (step two) or for contact tracing (step four) but instead is tobe used for a population study (step six), then only the minimum patientidentifying information in compliance with privacy regulations and/oragreements is transmitted and/or stored along with the received testreport (step seven). If the report is not for a trusted receiver,contact tracing or population study (step six) then it is determined ifthere is any legitimate use of the test report data and an action istaken accordingly or the automatically data is purged from storage.

FIG. 62 is a perspective view of an embodiment of a EBC/EBA collectionsystem. FIG. 63 is a perspective view of the EBC/EBA collection systemshowing a pipette and pipette guide. FIG. 64 is an exploded view showingthe constituent parts of the embodiment of the EBC/EBA collectionsystem. FIG. 65 is another exploded view showing the constituent partsof the EBC/EBA collection system. FIG. 66 is a cross-sectional view ofthe EBC/EBA collection system. FIG. 67 illustrates the use of theEBC/EBA collection system for obtaining biomarker samples from the lungsof a user.

In accordance with an aspect of the invention, an apparatus fordetecting a biomarker includes a particulate capturing structure forreceiving and capturing exhaled breath aerosol (EBA) particulate fromairway linings of a user, the particulate capturing structure having anaerosol particulate testing system for receiving the capturedparticulate and detecting a first biomarker, wherein the aerosolparticulate testing system includes a dissolvable EBA sample collectorfilm for capturing EBA particulate. The first reagent is bound to afirst nanoparticle and held in place at the insoluble testing area. TheEBA particulate includes non-soluble particulates and dropletparticulates, and the dissolvable EBA collector film includes a tackysurface for adhering to and capturing the non-soluble particulates andwater soluble bulk for capturing droplet particulates.

A droplet harvesting structure may be provided for converting breathvapor from a user to an exhaled breath condensate (EBC) fluid dropletfor forming a fluid sample. The user exhales through a mouthpiece sothat the exhaled breath impinges on the walls of an inner cylinder. Theinner cylinder can include a thermal mass (e.g., made for aluminum orother suitable material, or include an inner space that can be fulledwith a cold thermal mass). The walls of the inner cylinder receives thebreath vapor and forms the fluid droplet from the received breath vapor.The inner cylinder ends in a sharp point to help channel the fluiddroplet towards a sloped base. The sloped bass is the end of an outercylinder that collects that fluid sample from the inner walls of theouter cylinder and the outwalls of the inner cylinder. A pipette passesthrough a pipette hole in a cap and is used to draw the accumulatedfluid sample from the sloped base. Using the pipette, the user expelsdrops of the fluid sample onto a fluid sample testing system having abiomarker testing zone for receiving the fluid sample and detecting asecond biomarker. The cap may also include diverting structures to helpkeep the breath vapor in contact with the walls of the inner cylinder.All or parts of the system can be integrally formed from an injectionmold, or separate parts assembled into the completed system. The entiresystem or just the inner cylinder can be placed in a freezer ahead ofthe use to facilitate droplet collection from the chilled walls thatcome in contact with the breath vapor.

FIG. 68 is an isolated view showing the mouthpiece, cap, base,dissolvable EBA sample collector and inner cylinder of the embodiment ofthe EBC/EBA collection system. FIG. 69 is an isolated view showing thedissolvable EBA sample collector and inner cylinder having captured EBAparticles and droplets. FIG. 70 shows the inner cylinder submersed in asolvent for dissolving the dissolvable EBA sample collector to acquirethe captured EBA particles and droplets for biomarker testing.

FIG. 71 is an isolated view of a section of an embodiment of thedissolvable EBA sample collector forming an aerosol particulate testingsystem having captured EBA particulate, insoluble testing areas anddissolvable capture film areas. The dissolvable EBA sample collectorfilm includes a first reagent for reacting with at least one constituentof the captured particulate in a detection reaction for detecting thefirst biomarker. The detection reaction generates at least one of achange in an optical signal and an electrical signal dependent on thefirst biomarker. The detection reaction can in situ, so that with veryclosely spaced dissolvable and insoluble test areas, the EBA dropletsdissolve into the dissolvable film where an analyte in the droplet ispicked up, for example, by a labeled-antibody to form an analyte-labeledantibody complex that is bound to capture antibodies and retained at thenon-soluble test areas for visual or phontonics detection (similar tothe action of a Lateral Flow Assay as described herein). In this case,FIG. 72 shows a series of side views of the embodiment of thedissolvable EBS sample collector capturing EBA droplets and/orparticulate showing the aerosol particulate testing system with targetanalytes captured and bound to the insoluble testing areas.Alternatively, the captured EBA particulate and droplets can be sent infor analysis by a lab where a technician or automated system rinses thedissolvable film to provide a fluid sample that includes the capturedEBA analytes. For example, the inner cylinder can be rinsed with a flowor submersed in a solvent for dissolving the dissolvable EBA samplecollector to acquire the captured EBA particles and droplets forbiomarker testing.

FIG. 73 shows nanoparticles held in a trench in a substrate where thenanoparticles include capture antibodies or other reagent fixed to them.In this case, the fluid sample testing system may comprise a fluidicbiosensor for receiving the fluid sample potentially containing abiomarker analyte as the second biomarker and including a sample sourcehaving a biomarker analyte, a bioreceptor area functionalized with ananalyte-specific bioreceptor, and a transducer for generating a readablesignal depending on a change in the bioreceptor in response to receivingthe biomarker analyte from the sample source. The analyte-specificbiomarker includes a reagent for creating a detection reaction with thebiomarker analyte and where the fluidic biosensor generates at least oneof a change in an optical signal and an electrical signal dependent onthe biomarker. The reagent is bound to a nanoparticle and held in placeat the insoluble testing area.

FIG. 74 shows the EBA particles and droplets being rinsed from thedissolvable EBA sample collector to form a fluid sample that includesany biomarker analytes contained in the particles or droplets. This canbe done by the user using a solution that includes a buffer andsurfactant (and other materials, or these materials may be included inthe dissolvable film). This can also be done at a laboratory by atechnician or automated equipment.

FIG. 75 illustrates the EBA/EBC testing system with a wirelesscommunication electronic circuit that detects a result of the testingfor at least one of the first and second biomarker and communicating theresult to a wireless receiver. The wireless communication electroniccircuit in communication with at least one of the aerosol particulatetesting system and the fluid sample testing system for detecting one ormore biometric parameters, where the biometric parameters are dependenton at least one physiological change to a patient in response to aconcerning condition such as a virus infection where the one or morebiometric parameters are received and probabilistic analysis applied bya microprocessor to determine if at least one physiological changethreshold has been exceeded dependent on the probabilistic analysis ofthe one ore more biometric parameters and where the electronic circuittransmits a signal depending on the determined exceeded said at leastone physiological change.

In accordance with another aspect of the invention an apparatuscomprises at least one processor, at least one memory including computerprogram code, the at least one memory and the computer program codeconfigured to, with the at least one processor, cause the apparatus toperform at least the following: detecting one or more biometricparameters using a particulate capturing structure for receiving andcapturing exhaled breath aerosol (EBA) particulate from airway liningsof a user, the particulate capturing structure having an aerosolparticulate testing system for receiving the captured particulate anddetecting a first biomarker, wherein the aerosol particulate testingsystem includes a dissolvable EBA sample collector film for capturingEBA particulate, where the biometric parameters are biomarkers dependenton at least one physiological change to a patient in response to aconcerning condition such as a virus infection; receiving the one ormore biometric parameters and applying probabilistic analysis todetermine if at least one physiological change threshold has beenexceeded dependent on the probabilistic analysis of the one ore morebiometric parameters; and activating an action depending on thedetermined exceeded said at least one physiological change. The one ormore biometric parameters can be further detected using a dropletharvesting structure for converting breath vapor to a fluid droplet forforming a fluid sample and a testing system having a biomarker testingzone for receiving the fluid sample and detecting the biometricparameter; and wherein the probabilistic analysis is applied to the oneor more biometric parameters to determine if the at least onephysiological change threshold has been exceeded dependent on theprobabilistic analysis of the one ore more biometric parameters detectedfrom both the captured particulates and the fluid sample.

Various modifications and adaptations to the foregoing exemplaryembodiments of this invention may become apparent to those skilled inthe relevant arts in view of the foregoing description, when read inconjunction with the accompanying drawings. However, any and allmodifications will still fall within the scope of the non-limiting andexemplary embodiments of this invention.

The embodiments described herein are intended to exemplary andnon-limiting, the selection of biometric, environmental, or othermeasured conditions is not limited to a specific metric or multiplemetrics described herein but will depend on the particular applicationand treatment, data collection, and/or other use of the detectedmetrics. Also, the treatments employed in any of the embodimentsdescribed herein is not limited to a specific treatment or action butwill depend on the intended use and desired outcome of the combineddetected metrics and applied treatments.

Furthermore, some of the features of the various non-limiting andexemplary embodiments of this invention may be used to advantage withoutthe corresponding use of other features. As such, the foregoingdescription should be considered as merely illustrative of theprinciples, teachings and exemplary embodiments of this invention, andnot in limitation thereof. Various modifications and adaptations to theforegoing exemplary embodiments of this invention may become apparent tothose skilled in the relevant arts in view of the foregoing description,when read in conjunction with the accompanying drawings. However, anyand all modifications will still fall within the scope of thenon-limiting and exemplary embodiments of this invention.

The embodiments described herein are intended to exemplary andnon-limiting, the selection of biometric, environmental, or othermeasured conditions is not limited to a specific metric or multiplemetrics described herein but will depend on the particular applicationand treatment, data collection, and/or other use of the detectedmetrics. Also, the treatments employed in any of the embodimentsdescribed herein is not limited to a specific treatment or action butwill depend on the intended use and desired outcome of the combineddetected metrics and applied treatments.

Furthermore, some of the features of the various non-limiting andexemplary embodiments of this invention may be used to advantage withoutthe corresponding use of other features. As such, the foregoingdescription should be considered as merely illustrative of theprinciples, teachings and exemplary embodiments of this invention, andnot in limitation thereof.

What is claimed is:
 1. An apparatus for detecting a biomarker,comprising: a particulate capturing structure for receiving andcapturing exhaled breath aerosol (EBA) particulate from airway liningsof a user, the particulate capturing structure having an aerosolparticulate testing system for receiving the captured particulate anddetecting a first biomarker, wherein the aerosol particulate testingsystem includes a dissolvable EBA sample collector film for capturingEBA particulate.
 2. An apparatus for detecting a biomarker according toclaim 1; wherein the dissolvable EBA sample collector film includes afirst reagent for reacting with at least one constituent of the capturedparticulate in a detection reaction for detecting the first biomarker.3. An apparatus for detecting a biomarker according to claim 2; whereinthe detection reaction generates at least one of a change in an opticalsignal and an electrical signal dependent on the first biomarker.
 4. Anapparatus for detecting a biomarker according to claim 3; wherein thefirst reagent is bound to a first nanoparticle and held in place at theinsoluble testing area.
 5. An apparatus for detecting a biomarkeraccording to claim 1; wherein the EBA particulate includes non-solubleparticulates and droplet particulates, and the dissolvable EBA collectorfilm includes a tacky surface for adhering to and capturing thenon-soluble particulates and water soluble bulk for capturing dropletparticulates.
 6. An apparatus for detecting a biomarker according toclaim 1; further comprising a droplet harvesting structure forconverting breath vapor from a user to an exhaled breath condensate(EBC) fluid droplet for forming a fluid sample.
 7. An apparatus fordetecting a biomarker according to claim 6, wherein the dropletharvesting structure includes at least one of a hydrophobic field forreceiving the breath vapor and forming the fluid droplet from thereceived breath vapor and hydrophilic channels for receiving the fluiddroplet and channeling the fluid droplet towards a fluid sample testingsystem, the fluid sample testing system having a biomarker testing zonefor receiving the fluid sample and detecting a second biomarker.
 8. Anapparatus for detecting a biomarker according to claim 8, furthercomprising a fluid dam member disposed between the droplet harvestingstructure and the biomarker testing zone, wherein the fluid dam memberincludes at least one of a removable moisture resistant sheet member anda dissolvable film for accumulating the fluid sample from the dropletharvesting structure and releasing the accumulated fluid sample to flowto the biomarker testing zone.
 9. An apparatus for detecting a biomarkeraccord go to claim 8, wherein the fluid sample testing system comprisesa fluidic lateral flow assay including a sample pad for receiving thefluid sample potentially containing a biomarker analyte as the secondbiomarker, a conjugate release pad, a flow membrane and an adsorbent padfor receiving and flowing the fluid sample to detect the potentialbiomarker analyte from the sample source.
 10. An apparatus according toclaim 10, further comprising a fluid dam member disposed between thesample pad and the conjugate release pad, the fluid dam including a pulltab structure to enable a user to remove the fluid dam member and allowthe flow of the fluid sample from the sample pad to the conjugaterelease pad.
 11. An apparatus according to claim 8, further comprisingat least one photoemitter and one photodetector, wherein thephotoemitter emits radiation towards the biomarker testing zone and thephotodetector receives radiation from the biomarker testing zone.
 12. Anapparatus for detecting a biomarker according to claim 7 where the fluidsample testing system comprises a fluidic biosensor for receiving thefluid sample potentially containing a biomarker analyte as the secondbiomarker and including a sample source having a biomarker analyte, abioreceptor area functionalized with an analyte-specific bioreceptor,and a transducer for generating a readable signal depending on a changein the bioreceptor in response to receiving the biomarker analyte fromthe sample source.
 13. An apparatus for detecting a biomarker accordingto claim 12; where the analyte-specific biomarker includes a reagent forcreating a detection reaction with the biomarker analyte and where thefluidic biosensor generates at least one of a change in an opticalsignal and an electrical signal dependent on the biomarker.
 14. Anapparatus for detecting a biomarker according to claim 13; wherein thereagent is bound to a nanoparticle and held in place at the insolubletesting area.
 15. An apparatus according to claim 14; further comprisinga wireless communication electronic circuit for detecting a result ofthe testing for at least one of the first and second biomarker andcommunicating the result to a wireless receiver.
 16. A system accordingto claim 15; wherein the electronic circuit is in communication with atleast one of the aerosol particulate testing system and the fluid sampletesting system for detecting one or more biometric parameters, where thebiometric parameters are dependent on at least one physiological changeto a patient in response to a concerning condition such as a virusinfection where the one or more biometric parameters are received andprobabilistic analysis applied by a microprocessor to determine if atleast one physiological change threshold has been exceeded dependent onthe probabilistic analysis of the one ore more biometric parameters andwhere the electronic circuit transmits a signal depending on thedetermined exceeded said at least one physiological change.
 17. Anapparatus, comprising: at least one processor; and at least one memoryincluding computer program code, the at least one memory and thecomputer program code configured to, with the at least one processor,cause the apparatus to perform at least the following: detecting one ormore biometric parameters using a particulate capturing structure forreceiving and capturing exhaled breath aerosol (EBA) particulate fromairway linings of a user, the particulate capturing structure having anaerosol particulate testing system for receiving the capturedparticulate and detecting a first biomarker, wherein the aerosolparticulate testing system includes a dissolvable EBA sample collectorfilm for capturing EBA particulate, where the biometric parameters arebiomarkers dependent on at least one physiological change to a patientin response to a concerning condition such as a virus infection;receiving the one or more biometric parameters and applyingprobabilistic analysis to determine if at least one physiological changethreshold has been exceeded dependent on the probabilistic analysis ofthe one ore more biometric parameters; and activating an actiondepending on the determined exceeded said at least one physiologicalchange.
 18. An apparatus according to claim 17, wherein the one or morebiometric parameters are further detected using a droplet harvestingstructure for converting breath vapor to a fluid droplet for forming afluid sample and a testing system having a biomarker testing zone forreceiving the fluid sample and detecting the biometric parameter; andwherein the probabilistic analysis is applied to the one or morebiometric parameters to determine if the at least one physiologicalchange threshold has been exceeded dependent on the probabilisticanalysis of the one ore more biometric parameters detected from both thecaptured particulates and the fluid sample.